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ACT | Actin (polyclonal)

AS13 2640 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Agostis stoloniferacv. ‘Penncross’,Arabidopsis thaliana, Brassica sp., Cannabis sativa L., Cucumis sativus, Cynara cardunculus, Glycine max, Hordeum vulgare, Nicotiana tabacum, Phoenix dactylifera, Picrorhiza kurroa, Setaria italica, Solanum tuberosum, Triticum aestivum, Zea mays

ACT | Actin (polyclonal) in the group Antibodies Plant/Algal  / Plant Developmental Biology / Cytoskeleton at Agrisera AB (Antibodies for research) (AS13 2640)
ACT | Actin (polyclonal)



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Product Information

Immunogen

ca. 100 amino acids of recombinant actin conserved more than 80 %  in Arabidopsis thaliana: actin-1 P0CJ46  AT2G37620, actin-2 Q96292 AT3G18780, actin-3 P0CJ47 AT3G53750, actin-4 P53494 AT5G59370, actin-5 Q8RYC2 At2g42100, actin-7 P53492 At5g09810, actin-8 Q96293  AT1G49240 , actin-11 P53496 , AT3G12110 ,actin-12 P53497 AT3G46520

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution add 50 µl of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications immunofluorescence (IF), Western blot (WB)
Recommended dilution 1-100 - 1 : 250 (IF), 1 : 3000-1 : 5000 (WB)
Expected | apparent MW 41,6 | 45 kDa

Reactivity

Confirmed reactivity Agostis stolonifera cv. ‘Penncross’,Arabidopsis thaliana, Brassica napus, Cucumis sativus,Cyanthobasis fruticulosa, Cynara cardunculus, Glycine max, Hordeum vulgare, Nicotiana tabacum, Petrosimonia nigdeensis, Phoenix dactylifera, Picrorhiza kurroa,Salsola grandis, Salsola tragus, Setaria italica, Solanum tuberosum, Triticum aestivum, Zea mays, Vigna unguiculata
Predicted reactivity

Agropyron cristatum, Beta vulgaris, Betula luminifera, Brassica rapa subsp. pekinensis, Daucus carota, Cannabis sativa L. , Capsella rubella,Castanea sativa, Chorispora bungeana, Cyanidioschyzon merolae strain 10D,  Glycine soja, Halogeton glomeratus, Ipomoea batatas, Manihot esculenta, Medicago truncatula, Malus domestica, Oryza sativa, Pisum sativun, Populus sp., Solanum lycopersicum, Solanum tuberosum, Phaseolus vulgaris, Phaeodactylum tricornutum, Picea abies, Picea sitchensis, Prunus avium, Ricinus communis, Rubus plicatus, Theobroma cacao,Vicia faba


Species of your interest not listed? Contact us
Not reactive in

Chlamydomonas reinhardtii (too high background for this species)

Application examples

Application examples

Application example


western blot using anti-plant actin antibody

15 µg of total protein extracted with PEB (AS08 300) from  leaf tissue of (1) Arabidopsis thaliana, (2) Hordeum vulgare, (3) Zea mays were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-actin (AS13 2640, 1:2500, 1h) and secondary anti-rabbit (1:10 000, 1 h) antibody (HRP conjugated, recommended secondary antibody AS09 602) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection reagent using a Fuji LAS-3000 CCD (300s, standard sensitivity). Exposure time was 2 min.
 

 

 immunolocalization using anti-plant actin antibody

Actin cytoskeleton in 5 days old Arabidopsis thaliana seedlings. Actin signal shown in green, PIN1 in red and DAPI in blue. The material has been fixed in 2 % formaldehyde for 45 minutes. Tissue cleaning has been performed before immunolocalization. Rabbit anti-actin primary antibody was diluted in 1:250 and anti-rabbit Alexa 488 and Alexa 555 were both diluted in 1: 500 (Invitrogen). Scale bar - 20 µm.

Courtesy: Dr. Taras Pasternak, Freiburg University, Germany


 
Western blot of anti-actin antibodies on potato tuber flesh

Proteins were extracted from tuber flesh of Russet Burbank potato (Solanum tuberosum) with 0.1 M Tris HCl (pH=8.0), 5% sucrose (m/v), 2% (m/v) SDS, protease inhibitors (PMSF 1mM). Samples were heated 95°C 5 min, and 10 μg of total protein was resolved in 12% SDS PAGE and blotted to PVDF membrane for 1h-1.5h using tank transfer. Blots were blocked with a skimmed milk 4% (m/v) in T-TBS (1.5h) at RT with agitation. Primary antibodies (AS13 2640) were applied overnight +4°C in dilution 1:5000 with agitation. After washing with T-TBS 2-3 times, membrane was incubated with secondary antibodies (Goat Anti-Rabbit HRP conjugate, Transgen biotech HS101) 1:10000 for 1 hour at RT. Blot was washed as above and developed with ECL (Clarity Western ECL Substrate, BioRad, 170-5060) for 5 – 10 minutes. Exposure time – 20.395 seconds.

Courtesy of Iauhenia Isayenka, University of Sherbrooke, Canada


Reactant: Solanum lycopersicum (Tomato)

Application: Western Blotting

Pudmed ID: 26347766

Journal: Front Plant Sci

Figure Number: 6A

Published Date: 2015-09-09

First Author: Buxa, S. V., Degola, F., et al.

Impact Factor: 5.435

Open Publication

(A–D) Western blot analyses of S. lycopersicum midrib extracts. Three healthy (lanes 1–3) and three infected (lanes 4–6) plants were analyzed. Immunoreactive signals corresponding to actin and ER luminal binding protein (BiP) displayed the expected molecular weights of ?40 and ?78 kDa, respectively (A). Amount of loaded protein were checked by Ponceau-S staining (B). Quantification of immunoreactive luminescence signals was achieved by densitometric scanning of the respective actin (C) and BiP (D) bands. The mean of six replicates (±SD) was calculated for each sample. Actin T-value: –9.977; BiP T-value: 15.068. * denotes significant difference of the means (P-values ? 0.001).


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 27406783

Journal: J Exp Bot

Figure Number: 7A

Published Date: 2016-09-01

First Author: Sztatelman, O., ?abuz, J., et al.

Impact Factor: 6.088

Open Publication

Representative dephosphorylation profiles of phototropin1 after blue light exposure (120 µmol m?2 s?1 and 1h) in Arabidopsis wild-type and mutant (phot2 and rcn1) leaves. Dark, a dark-adapted control; 0, a sample collected just after illumination. The duration of the incubation in the darkness after the end of the illumination is indicated in minutes. Phosphorylation leads to the shift of the phototropin band towards higher mass. Samples treated with alkaline phosphatase are shown on the right. Anti-actin blots are presented as the loading reference. The results represent two out of 4–5 independent biological replicates.


Reactant: Ralstonia solanacearum

Application: Western Blotting

Pudmed ID: 29265643

Journal: Plant Biotechnol J

Figure Number: 4A

Published Date: 2018-07-01

First Author: Wei, Y., Caceres-Moreno, C., et al.

Impact Factor: 8.66

Open Publication

Csp22 from Ralstonia solanacearum triggers additional early and late PRR?dependent immune responses in Solanaceae plants. (a) MAPK activation assay in leaves of 3? or 5?week?old Nicotiana benthamiana plants after treatment with 1 ?m csp22Rsol or water (mock) for 15 or 30 min as indicated. Immunoblots were analysed using antiphosphorylated MAPK antibody (??pMAPK). Immunoblots were also analysed using ??actin antibody to verify protein accumulation. Molecular weight (kDa) marker bands are indicated for reference. (b) Growth inhibition of N. benthamiana seedlings treated with 100 nm flg22Pto, csp22Rsol or water (mock) for 9 days. Values represent seedling weigh shown as percentage of mock?treated seedlings (average ± SE; n = 12). (c) Photograph of seedlings after exposure to the different elicitors in (b). (d) Growth of R. solanacearum GMI1000 in tomato (cultivar M82) leaves pretreated with 1 ?m csp22Rsol or water (mock) for 24 h and then syringe?infiltrated with a 106 cfu/mL bacterial inoculum. Bacterial growth was determined 3 days after inoculation. Values are mean ± SE (n = 3). Asterisks indicate significant differences compared to the mock control according to a Student's t?test (P < 0.001). All experiments were performed three times with similar results.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 29365132

Journal: J Exp Bot

Figure Number: 1B

Published Date: 2018-03-14

First Author: Minina, E. A., Moschou, P. N., et al.

Impact Factor: 6.088

Open Publication

Constitutive overexpression of ATG5 or ATG7 in Arabidopsis stimulates lipidation of Atg8. (A) A schematic representation of two autophagy-related ubiquitin-like conjugation systems (Nakatogawa, 2013). Atg7 acts as an E1-like ligase by stimulating conjugation of two substrate pairs: of a ubiquitin-like protein Atg12 with its E2-like ligase, Atg10, and of a ubiquitin-like protein Atg8 with its E2-like ligase, Atg3. Atg12 is further transferred from Atg10 onto Atg5 protein in an E3-ligase-independent manner. The Atg12–Atg5 conjugate forms a complex with Atg16 and gains an E3-like ligase activity. Atg12–Atg5–Atg16 E3-like ligase stimulates conjugation of Atg8 with a phosphatidylethanolamine (PE), followed by anchoring of Atg8–PE on a membrane of a growing phagophore. Double green and double red asterisks indicate Atg12–Atg5 and Atg8–PE conjugates, respectively, also shown on the western blots on (B) and (D). (B) Atg5, but not Atg7, is a limiting component in the Atg12–Atg5 conjugation pathway. Western blot detection of Atg5 in rosette leaves of Col-0 (WT), two individual ATG5- or ATG7-overexpressing lines (ATG5 OE or ATG7 OE, respectively), and ATG5- or ATG7-knockout (atg5 or atg7, respectively) mutants. *Atg5; **Atg12–Atg5 conjugate. The experiment was repeated four times, using three individual lines of each overexpressor background. Western blot detection of actin and Ponceau staining were used as loading controls. (C) Densitometry of the Atg5–Atg12 conjugate in (B). Integrated density of bands corresponding to the Atg5–Atg12 conjugate were first normalized to the corresponding values for actin and then expressed as a percentage of the obtained value for the WT. Data represent means ±SE; n=6. Mean values denoted by the same letter do not differ significantly at P<0.001 (Student’s t-test). (D) Up-regulation of either ATG5 or ATG7 stimulates lipidation of Atg8. Seven-day-old seedlings of Col-0 (WT), atg5, ATG5 OE, atg7, and ATG7 OE genotypes were incubated for 3 d under 150 µE m–2 s–1 light, 16 h photoperiod (Light) or in the darkness (Dark). Total proteins were fractionated by ultracentrifugation and used for western blot analysis to detect free (*) and lipidated (**) forms of Atg8. C, crude extract; S, supernatant of 100000 g fraction; M, pellet of 100000 g fraction, containing membranes. (E) Densitometry of the Atg8–PE conjugate in (D). Integrated density of bands corresponding to the Atg8–PE in the crude fraction was expressed as a percentage of the integrated density of the total amount of Atg8 in the corresponding sample. Data represent means ±SE; n=6. Mean values denoted by the same letter do not differ significantly at P<0.001 (Student’s t-test). L, light; D, dark.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 29365132

Journal: J Exp Bot

Figure Number: 2C

Published Date: 2018-03-14

First Author: Minina, E. A., Moschou, P. N., et al.

Impact Factor: 6.088

Open Publication

Constitutive overexpression of ATG5 or ATG7 stimulates formation of autophagosomes and increases autophagic flux. (A) Representative confocal microscopy images of the epidermal root cells expressing GFP–Atg8a in WT, ATG-knockout, and ATG-overexpressing backgrounds. Seedlings were grown under 150 µE m–2 s–1 light, 16 h photoperiod for 6 d and subjected to 0.5 µM ConA treatment for 16 h prior to imaging. Scale bars=10 µm. (B) The box-plot diagram shows the number of the GFP-positive puncta in the epidermal root cells in (A). The average number of puncta for two or three cells belonging to the same root was considered as a single measurement. Four to seven roots were imaged per genotype. The means of each genotype were compared with the WT using Dunnett’s test, assuming a Poisson distribution of the data. ***P<0.001; ?, outliers. (C) Increase of autophagic flux in ATG5- or ATG7-overexpressing plants confirmed by a higher rate of NBR1 degradation. Plants of the indicated genotypes were grown under 150 µE m–2 s–1. Rosette leaves were sampled at the onset of flowering (0 days after flowering, DAF) and 10 d later (10 DAF). Total protein extracts from sampled leaves were used for western blot analysis to detect NBR1 and actin. (D) Densitometry of the NBR1 signal in (C) and expression level of NBR1 in the same samples. Integrated density values for NBR1 were first normalized to actin and then expressed as a percentage of 0 DAF for the corresponding sample. Data represent means ±SE; n=6. The means of each genotype were compared with the WT using Dunnett’s test, ***P<0.001. Detection of the NBR1 transcript level in ATG-overexpressing plants confirmed that the decrease in NBR1 protein was caused by protein degradation and did not originate from transcriptional variation. qRT-PCR was performed on the same leaf material as in (C). Data represent means ±SE for each genotype, normalized to two reference genes (PP2A and HEL) and to 0 DAF; n=6. The means of each genotype were compared with the WT using Dunnett’s test, ***P<0.001. (E) Increased autophagic flux in ATG5- or ATG7-overexpressing plants additionally confirmed by detection of free GFP accumulation in the seedlings expressing GFP–Atg8a in the indicated backgrounds. Seven-day-old seedlings grown under normal conditions (150 µE m–2 s–1 light, 16 h photoperiod) were either transferred onto sucrose-depleted MS medium and kept in the darkness (–) or left growing under normal conditions (+). Total protein extracts from the whole seedlings of each genotype were used for western blot analysis with anti-GFP. The GFP–Atg8a fusion has a predicted molecular weight of ~40 kDa; free GFP has a predicted molecular weight of 27 kDa. To avoid quantification of saturated pixels, several exposures were used for different samples. Comparisons of the absolute integrated density values for each line were made using the same exposure. For more information, see the Materials and methods. (F) Densitometry of the GFP–ATG8a cleavage assay in (E) confirms elevated autophagic flux in ATG OE backgrounds. The experiment was repeated twice; at least three western blot assays were performed for each experiment. The figure shows a representative example. Free GFP was expressed as a percentage of the total amount of GFP for each sample in (E). Data represent means ±SE of three individual measurements. The means of each genotype were compared with the WT using Dunnett’s test, ***P<0.001.

Additional information

Additional information Antibody available in 3 various pack sizes: 50, 100 and 150 µl - Please inquire.

This product can be sold containing ProClin if requested.

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Background

Background

Actin is a highly conserved protein and an essential component of cell cytoskeleton and plays an important role in cytoplasmic streaming, cell shape determination, cell division, organelle movement and extension growth. Preferentially expressed in young and expanding tissues, floral organ primordia, developing seeds and emerging inflorescence.

Product citations

Selected references Czernicka et al. (2022). Proteomic Studies of Roots in Hypoxia-Sensitive and -Tolerant Tomato Accessions Reveal Candidate Proteins Associated with Stress Priming. Cells. 2022 Jan 31;11(3):500. doi: 10.3390/cells11030500. PMID: 35159309; PMCID: PMC8834170.
Kumari et al. (2021) In-depth assembly of organ and development dissected Picrorhiza kurroa proteome map using mass spectrometry. BMC Plant Biol. 2021 Dec 22;21(1):604. doi: 10.1186/s12870-021-03394-8. PMID: 34937558; PMCID: PMC8693493.
Mishra, Sahu & Shaw (2021). Insight into the cellular and physiological regulatory modulations of Class-I TCP9 to enhance drought and salinity stress tolerance in cowpea. Physiol Plant. 2021 Aug 30. doi: 10.1111/ppl.13542. Epub ahead of print. PMID: 34459503.
Zhuang et al (2021). EGY3 mediates chloroplastic ROS homeostasis and promotes retrograde signaling in response to salt stress in Arabidopsis. Cell Rep. 2021 Jul 13;36(2):109384. doi: 10.1016/j.celrep.2021.109384. PMID: 34260941.
Ngou et al. (2021) Mutual potentiation of plant immunity by cell-surface and intracellular receptors. Nature. 2021 Mar 10. doi: 10.1038/s41586-021-03315-7. Epub ahead of print. PMID: 33692545.
labuz et al. (2021) Phototropin interactions with SUMO proteins. Plant Cell Physiol. 2021 Feb 17:pcab027. doi: 10.1093/pcp/pcab027. Epub ahead of print. PMID: 33594440.
Khajuria et al. (2020). Photochemical Efficiency Is Negatively Correlated With the Delta 9- Tetrahydrocannabinol Content in Cannabis Sativa L. Plant Physiol Biochem. 2020 Apr 8;151:589-600. doi: 10.1016/j.plaphy.2020.04.003.
Molnar et al. (2020). Nitro-oxidative Signalling Induced by Chemically Synthetized Zinc Oxide Nanoparticles (ZnO NPs) in Brassica Species. Chemosphere, 251, 126419
Roustan et al. (2020). Protein sorting into protein bodies during barley endosperm development is putatively regulated by cytoskeleton members, MVBs and the HvSNF7s. Sci Rep. 2020 Feb 5;10(1):1864. doi: 10.1038/s41598-020-58740-x.
Dalmadi et al. (2019). AGO-unbound cytosolic pool of mature miRNAs in plant cells reveals a novel regulatory step at AGO1 loading. Nucleic Acids Res. 2019 Aug 8. pii: gkz690. doi: 10.1093/nar/gkz690.
Patankar et al. (2019). Functional Characterization of Date Palm Aquaporin Gene PdPIP1;2 Confers Drought and Salinity Tolerance to Yeast and Arabidopsis. Genes (Basel). 2019 May 22;10(5). pii: E390. doi: 10.3390/genes10050390.
Scherer et al. (2019). Pulsed electric field (PEF)-assisted protein recovery from Chlorella vulgaris is mediated by an enzymatic process after cell death. Algal Research, Volume 41, August 2019, 101536.
Czobor et al. (2019). Comparison of the response of alternative oxidase and uncoupling proteins to bacterial elicitor induced oxidative burst. PLoS One. 2019 Jan 10;14(1):e0210592. doi: 10.1371/journal.pone.0210592.
Deng et al. (2019). Integrated proteome analyses of wheat glume and awn reveal central drought response proteins under water deficit conditions. J Plant Physiol. 2019 Jan;232:270-283. doi: 10.1016/j.jplph.2018.11.011.
Zhang et al. (2018) DELLA proteins negatively regulate dark-induced senescence and chlorophyll degradation in Arabidopsis through interaction with the transcription factor WRKY6. Plant Cell Rep. 2018 Jul;37(7):981-992. doi: 10.1007/s00299-018-2282-9. Epub 2018 Mar 24. PMID: 29574486.
Wang et al. (2018). A role of GUNs-Involved retrograde signaling in regulating Acetyl-CoA carboxylase 2 in Arabidopsis. Biochem Biophys Res Commun. 2018 Nov 2;505(3):712-719. doi: 10.1016/j.bbrc.2018.09.144.
Borovik and Grabelnych (2018). Mitochondrial alternative cyanide-resistant oxidase is involved in an increase of heat stress tolerance in spring wheat. J Plant Physiol. 2018 Dec;231:310-317. doi: 10.1016/j.jplph.2018.10.007.
Pan et al. (2018). Comparative proteomic investigation of drought responses in foxtail millet. BMC Plant Biol. 2018 Nov 29;18(1):315. doi: 10.1186/s12870-018-1533-9.
Lopez?Calcagno et al. (2018). Overexpressing the H?protein of the glycine cleavage system increases biomass yield in glasshouse and field?grown transgenic tobacco plants. Plant Biotechnology Journal, May 2018.
Li et al. (2018). Comparative proteomic analysis of key proteins during abscisic acid-hydrogen peroxide-induced adventitious rooting in cucumber (Cucumis sativus L.) under drought stress. Journal of Plant Physiology Volume 229, October 2018, Pages 185-194.
Adhikari et al. (2018). Sulfate improves cadmium tolerance by limiting cadmium accumulation, modulation of sulfur metabolism and antioxidant defense system in maize. Environmental and Experimental Botany Volume 153, September 2018, Pages 143-162.
Brandt et al. (2018). Extended darkness induces internal turnover of glucosinolates in Arabidopsis thaliana leaves. PLoS One. 2018 Aug 9;13(8):e0202153. doi: 10.1371/journal.pone.0202153.
Jespersen et al. (2017). Metabolic Effects of Acibenzolar-S-Methyl for Improving Heat or Drought Stress in Creeping Bentgrass. Front Plant Sci. 2017 Jul 11;8:1224. doi: 10.3389/fpls.2017.01224. eCollection 2017. (western blot, Agostis stolonifera cv. ‘Penncross’)
Qiu et al. (2015). Soy 14-3-3 protein SGF14c, a new regulator of tolerance to salt–alkali stress. Plant Biotechnology Reports pp 1-9.
Shaw et al. (2015). Beta-aminobutyric acid mediated drought stress alleviation in maize (Zea mays L.). Environ Sci Pollut Res Int. 2015 Sep 29.
Buxa et al. (2015). Phytoplasma infection in tomato is associated with re-organization of plasma membrane, ER stacks, and actin filaments in sieve elements. ront Plant Sci. 2015; 6: 650. Published online 2015 Aug 19.
Zheng et al. (2014). iTRAQ-based quantitative proteomics analysis revealed alterations of carbohydrate metabolism pathways and mitochondrial proteins in a male sterile cybrid pummelo. J Proteome Res. 2014 May 13.
immunogen:

ca. 100 amino acids of recombinant actin conserved more than 80 %  in Arabidopsis thaliana: actin-1 P0CJ46  AT2G37620, actin-2 Q96292 AT3G18780, actin-3 P0CJ47 AT3G53750, actin-4 P53494 AT5G59370, actin-5 Q8RYC2 At2g42100, actin-7 P53492 At5g09810, actin-8 Q96293  AT1G49240 , actin-11 P53496 , AT3G12110 ,actin-12 P53497 AT3G46520

Reconstitution: For reconstitution add 50 µl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 50 µl
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: immunofluorescence (IF), Western blot (WB)
recommended dilution: 1-100 - 1 : 250 (IF), 1 : 3000-1 : 5000 (WB)
calculated | apparent molecular mass [kDa]: 41,6 | 45 kDa
Confirmed reactivity: Agostis stolonifera cv. ‘Penncross’,Arabidopsis thaliana, Brassica napus, Cucumis sativus,Cyanthobasis fruticulosa, Cynara cardunculus, Glycine max, Hordeum vulgare, Nicotiana tabacum, Petrosimonia nigdeensis, Phoenix dactylifera, Picrorhiza kurroa,Salsola grandis, Salsola tragus, Setaria italica, Solanum tuberosum, Triticum aestivum, Zea mays, Vigna unguiculata
predicted reactivity:

Agropyron cristatum, Beta vulgaris, Betula luminifera, Brassica rapa subsp. pekinensis, Daucus carota, Cannabis sativa L. , Capsella rubella,Castanea sativa, Chorispora bungeana, Cyanidioschyzon merolae strain 10D,  Glycine soja, Halogeton glomeratus, Ipomoea batatas, Manihot esculenta, Medicago truncatula, Malus domestica, Oryza sativa, Pisum sativun, Populus sp., Solanum lycopersicum, Solanum tuberosum, Phaseolus vulgaris, Phaeodactylum tricornutum, Picea abies, Picea sitchensis, Prunus avium, Ricinus communis, Rubus plicatus, Theobroma cacao,Vicia faba


Species of your interest not listed? Contact us
not reactive in:

Chlamydomonas reinhardtii (too high background for this species)

Picture (footer):

Application example


western blot using anti-plant actin antibody

15 µg of total protein extracted with PEB (AS08 300) from  leaf tissue of (1) Arabidopsis thaliana, (2) Hordeum vulgare, (3) Zea mays were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-actin (AS13 2640, 1:2500, 1h) and secondary anti-rabbit (1:10 000, 1 h) antibody (HRP conjugated, recommended secondary antibody AS09 602) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection reagent using a Fuji LAS-3000 CCD (300s, standard sensitivity). Exposure time was 2 min.
 

 

 immunolocalization using anti-plant actin antibody

Actin cytoskeleton in 5 days old Arabidopsis thaliana seedlings. Actin signal shown in green, PIN1 in red and DAPI in blue. The material has been fixed in 2 % formaldehyde for 45 minutes. Tissue cleaning has been performed before immunolocalization. Rabbit anti-actin primary antibody was diluted in 1:250 and anti-rabbit Alexa 488 and Alexa 555 were both diluted in 1: 500 (Invitrogen). Scale bar - 20 µm.

Courtesy: Dr. Taras Pasternak, Freiburg University, Germany


 
Western blot of anti-actin antibodies on potato tuber flesh

Proteins were extracted from tuber flesh of Russet Burbank potato (Solanum tuberosum) with 0.1 M Tris HCl (pH=8.0), 5% sucrose (m/v), 2% (m/v) SDS, protease inhibitors (PMSF 1mM). Samples were heated 95°C 5 min, and 10 μg of total protein was resolved in 12% SDS PAGE and blotted to PVDF membrane for 1h-1.5h using tank transfer. Blots were blocked with a skimmed milk 4% (m/v) in T-TBS (1.5h) at RT with agitation. Primary antibodies (AS13 2640) were applied overnight +4°C in dilution 1:5000 with agitation. After washing with T-TBS 2-3 times, membrane was incubated with secondary antibodies (Goat Anti-Rabbit HRP conjugate, Transgen biotech HS101) 1:10000 for 1 hour at RT. Blot was washed as above and developed with ECL (Clarity Western ECL Substrate, BioRad, 170-5060) for 5 – 10 minutes. Exposure time – 20.395 seconds.

Courtesy of Iauhenia Isayenka, University of Sherbrooke, Canada
More images:


Reactant: Solanum lycopersicum (Tomato)

Application: Western Blotting

Pudmed ID: 26347766

Journal: Front Plant Sci

Figure Number: 6A

Published Date: 2015-09-09

First Author: Buxa, S. V., Degola, F., et al.

Impact Factor: 5.435

Open Publication

(A–D) Western blot analyses of S. lycopersicum midrib extracts. Three healthy (lanes 1–3) and three infected (lanes 4–6) plants were analyzed. Immunoreactive signals corresponding to actin and ER luminal binding protein (BiP) displayed the expected molecular weights of ?40 and ?78 kDa, respectively (A). Amount of loaded protein were checked by Ponceau-S staining (B). Quantification of immunoreactive luminescence signals was achieved by densitometric scanning of the respective actin (C) and BiP (D) bands. The mean of six replicates (±SD) was calculated for each sample. Actin T-value: –9.977; BiP T-value: 15.068. * denotes significant difference of the means (P-values ? 0.001).


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 27406783

Journal: J Exp Bot

Figure Number: 7A

Published Date: 2016-09-01

First Author: Sztatelman, O., ?abuz, J., et al.

Impact Factor: 6.088

Open Publication

Representative dephosphorylation profiles of phototropin1 after blue light exposure (120 µmol m?2 s?1 and 1h) in Arabidopsis wild-type and mutant (phot2 and rcn1) leaves. Dark, a dark-adapted control; 0, a sample collected just after illumination. The duration of the incubation in the darkness after the end of the illumination is indicated in minutes. Phosphorylation leads to the shift of the phototropin band towards higher mass. Samples treated with alkaline phosphatase are shown on the right. Anti-actin blots are presented as the loading reference. The results represent two out of 4–5 independent biological replicates.


Reactant: Ralstonia solanacearum

Application: Western Blotting

Pudmed ID: 29265643

Journal: Plant Biotechnol J

Figure Number: 4A

Published Date: 2018-07-01

First Author: Wei, Y., Caceres-Moreno, C., et al.

Impact Factor: 8.66

Open Publication

Csp22 from Ralstonia solanacearum triggers additional early and late PRR?dependent immune responses in Solanaceae plants. (a) MAPK activation assay in leaves of 3? or 5?week?old Nicotiana benthamiana plants after treatment with 1 ?m csp22Rsol or water (mock) for 15 or 30 min as indicated. Immunoblots were analysed using antiphosphorylated MAPK antibody (??pMAPK). Immunoblots were also analysed using ??actin antibody to verify protein accumulation. Molecular weight (kDa) marker bands are indicated for reference. (b) Growth inhibition of N. benthamiana seedlings treated with 100 nm flg22Pto, csp22Rsol or water (mock) for 9 days. Values represent seedling weigh shown as percentage of mock?treated seedlings (average ± SE; n = 12). (c) Photograph of seedlings after exposure to the different elicitors in (b). (d) Growth of R. solanacearum GMI1000 in tomato (cultivar M82) leaves pretreated with 1 ?m csp22Rsol or water (mock) for 24 h and then syringe?infiltrated with a 106 cfu/mL bacterial inoculum. Bacterial growth was determined 3 days after inoculation. Values are mean ± SE (n = 3). Asterisks indicate significant differences compared to the mock control according to a Student's t?test (P < 0.001). All experiments were performed three times with similar results.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 29365132

Journal: J Exp Bot

Figure Number: 1B

Published Date: 2018-03-14

First Author: Minina, E. A., Moschou, P. N., et al.

Impact Factor: 6.088

Open Publication

Constitutive overexpression of ATG5 or ATG7 in Arabidopsis stimulates lipidation of Atg8. (A) A schematic representation of two autophagy-related ubiquitin-like conjugation systems (Nakatogawa, 2013). Atg7 acts as an E1-like ligase by stimulating conjugation of two substrate pairs: of a ubiquitin-like protein Atg12 with its E2-like ligase, Atg10, and of a ubiquitin-like protein Atg8 with its E2-like ligase, Atg3. Atg12 is further transferred from Atg10 onto Atg5 protein in an E3-ligase-independent manner. The Atg12–Atg5 conjugate forms a complex with Atg16 and gains an E3-like ligase activity. Atg12–Atg5–Atg16 E3-like ligase stimulates conjugation of Atg8 with a phosphatidylethanolamine (PE), followed by anchoring of Atg8–PE on a membrane of a growing phagophore. Double green and double red asterisks indicate Atg12–Atg5 and Atg8–PE conjugates, respectively, also shown on the western blots on (B) and (D). (B) Atg5, but not Atg7, is a limiting component in the Atg12–Atg5 conjugation pathway. Western blot detection of Atg5 in rosette leaves of Col-0 (WT), two individual ATG5- or ATG7-overexpressing lines (ATG5 OE or ATG7 OE, respectively), and ATG5- or ATG7-knockout (atg5 or atg7, respectively) mutants. *Atg5; **Atg12–Atg5 conjugate. The experiment was repeated four times, using three individual lines of each overexpressor background. Western blot detection of actin and Ponceau staining were used as loading controls. (C) Densitometry of the Atg5–Atg12 conjugate in (B). Integrated density of bands corresponding to the Atg5–Atg12 conjugate were first normalized to the corresponding values for actin and then expressed as a percentage of the obtained value for the WT. Data represent means ±SE; n=6. Mean values denoted by the same letter do not differ significantly at P<0.001 (Student’s t-test). (D) Up-regulation of either ATG5 or ATG7 stimulates lipidation of Atg8. Seven-day-old seedlings of Col-0 (WT), atg5, ATG5 OE, atg7, and ATG7 OE genotypes were incubated for 3 d under 150 µE m–2 s–1 light, 16 h photoperiod (Light) or in the darkness (Dark). Total proteins were fractionated by ultracentrifugation and used for western blot analysis to detect free (*) and lipidated (**) forms of Atg8. C, crude extract; S, supernatant of 100000 g fraction; M, pellet of 100000 g fraction, containing membranes. (E) Densitometry of the Atg8–PE conjugate in (D). Integrated density of bands corresponding to the Atg8–PE in the crude fraction was expressed as a percentage of the integrated density of the total amount of Atg8 in the corresponding sample. Data represent means ±SE; n=6. Mean values denoted by the same letter do not differ significantly at P<0.001 (Student’s t-test). L, light; D, dark.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 29365132

Journal: J Exp Bot

Figure Number: 2C

Published Date: 2018-03-14

First Author: Minina, E. A., Moschou, P. N., et al.

Impact Factor: 6.088

Open Publication

Constitutive overexpression of ATG5 or ATG7 stimulates formation of autophagosomes and increases autophagic flux. (A) Representative confocal microscopy images of the epidermal root cells expressing GFP–Atg8a in WT, ATG-knockout, and ATG-overexpressing backgrounds. Seedlings were grown under 150 µE m–2 s–1 light, 16 h photoperiod for 6 d and subjected to 0.5 µM ConA treatment for 16 h prior to imaging. Scale bars=10 µm. (B) The box-plot diagram shows the number of the GFP-positive puncta in the epidermal root cells in (A). The average number of puncta for two or three cells belonging to the same root was considered as a single measurement. Four to seven roots were imaged per genotype. The means of each genotype were compared with the WT using Dunnett’s test, assuming a Poisson distribution of the data. ***P<0.001; ?, outliers. (C) Increase of autophagic flux in ATG5- or ATG7-overexpressing plants confirmed by a higher rate of NBR1 degradation. Plants of the indicated genotypes were grown under 150 µE m–2 s–1. Rosette leaves were sampled at the onset of flowering (0 days after flowering, DAF) and 10 d later (10 DAF). Total protein extracts from sampled leaves were used for western blot analysis to detect NBR1 and actin. (D) Densitometry of the NBR1 signal in (C) and expression level of NBR1 in the same samples. Integrated density values for NBR1 were first normalized to actin and then expressed as a percentage of 0 DAF for the corresponding sample. Data represent means ±SE; n=6. The means of each genotype were compared with the WT using Dunnett’s test, ***P<0.001. Detection of the NBR1 transcript level in ATG-overexpressing plants confirmed that the decrease in NBR1 protein was caused by protein degradation and did not originate from transcriptional variation. qRT-PCR was performed on the same leaf material as in (C). Data represent means ±SE for each genotype, normalized to two reference genes (PP2A and HEL) and to 0 DAF; n=6. The means of each genotype were compared with the WT using Dunnett’s test, ***P<0.001. (E) Increased autophagic flux in ATG5- or ATG7-overexpressing plants additionally confirmed by detection of free GFP accumulation in the seedlings expressing GFP–Atg8a in the indicated backgrounds. Seven-day-old seedlings grown under normal conditions (150 µE m–2 s–1 light, 16 h photoperiod) were either transferred onto sucrose-depleted MS medium and kept in the darkness (–) or left growing under normal conditions (+). Total protein extracts from the whole seedlings of each genotype were used for western blot analysis with anti-GFP. The GFP–Atg8a fusion has a predicted molecular weight of ~40 kDa; free GFP has a predicted molecular weight of 27 kDa. To avoid quantification of saturated pixels, several exposures were used for different samples. Comparisons of the absolute integrated density values for each line were made using the same exposure. For more information, see the Materials and methods. (F) Densitometry of the GFP–ATG8a cleavage assay in (E) confirms elevated autophagic flux in ATG OE backgrounds. The experiment was repeated twice; at least three western blot assays were performed for each experiment. The figure shows a representative example. Free GFP was expressed as a percentage of the total amount of GFP for each sample in (E). Data represent means ±SE of three individual measurements. The means of each genotype were compared with the WT using Dunnett’s test, ***P<0.001.

additional information: Antibody available in 3 various pack sizes: 50, 100 and 150 µl - Please inquire.

This product can be sold containing ProClin if requested.

background:

Actin is a highly conserved protein and an essential component of cell cytoskeleton and plays an important role in cytoplasmic streaming, cell shape determination, cell division, organelle movement and extension growth. Preferentially expressed in young and expanding tissues, floral organ primordia, developing seeds and emerging inflorescence.

All references: Czernicka et al. (2022). Proteomic Studies of Roots in Hypoxia-Sensitive and -Tolerant Tomato Accessions Reveal Candidate Proteins Associated with Stress Priming. Cells. 2022 Jan 31;11(3):500. doi: 10.3390/cells11030500. PMID: 35159309; PMCID: PMC8834170.
Kumari et al. (2021) In-depth assembly of organ and development dissected Picrorhiza kurroa proteome map using mass spectrometry. BMC Plant Biol. 2021 Dec 22;21(1):604. doi: 10.1186/s12870-021-03394-8. PMID: 34937558; PMCID: PMC8693493.
Mishra, Sahu & Shaw (2021). Insight into the cellular and physiological regulatory modulations of Class-I TCP9 to enhance drought and salinity stress tolerance in cowpea. Physiol Plant. 2021 Aug 30. doi: 10.1111/ppl.13542. Epub ahead of print. PMID: 34459503.
Zhuang et al (2021). EGY3 mediates chloroplastic ROS homeostasis and promotes retrograde signaling in response to salt stress in Arabidopsis. Cell Rep. 2021 Jul 13;36(2):109384. doi: 10.1016/j.celrep.2021.109384. PMID: 34260941.
Ngou et al. (2021) Mutual potentiation of plant immunity by cell-surface and intracellular receptors. Nature. 2021 Mar 10. doi: 10.1038/s41586-021-03315-7. Epub ahead of print. PMID: 33692545.
labuz et al. (2021) Phototropin interactions with SUMO proteins. Plant Cell Physiol. 2021 Feb 17:pcab027. doi: 10.1093/pcp/pcab027. Epub ahead of print. PMID: 33594440.
Khajuria et al. (2020). Photochemical Efficiency Is Negatively Correlated With the Delta 9- Tetrahydrocannabinol Content in Cannabis Sativa L. Plant Physiol Biochem. 2020 Apr 8;151:589-600. doi: 10.1016/j.plaphy.2020.04.003.
Molnar et al. (2020). Nitro-oxidative Signalling Induced by Chemically Synthetized Zinc Oxide Nanoparticles (ZnO NPs) in Brassica Species. Chemosphere, 251, 126419
Roustan et al. (2020). Protein sorting into protein bodies during barley endosperm development is putatively regulated by cytoskeleton members, MVBs and the HvSNF7s. Sci Rep. 2020 Feb 5;10(1):1864. doi: 10.1038/s41598-020-58740-x.
Dalmadi et al. (2019). AGO-unbound cytosolic pool of mature miRNAs in plant cells reveals a novel regulatory step at AGO1 loading. Nucleic Acids Res. 2019 Aug 8. pii: gkz690. doi: 10.1093/nar/gkz690.
Patankar et al. (2019). Functional Characterization of Date Palm Aquaporin Gene PdPIP1;2 Confers Drought and Salinity Tolerance to Yeast and Arabidopsis. Genes (Basel). 2019 May 22;10(5). pii: E390. doi: 10.3390/genes10050390.
Scherer et al. (2019). Pulsed electric field (PEF)-assisted protein recovery from Chlorella vulgaris is mediated by an enzymatic process after cell death. Algal Research, Volume 41, August 2019, 101536.
Czobor et al. (2019). Comparison of the response of alternative oxidase and uncoupling proteins to bacterial elicitor induced oxidative burst. PLoS One. 2019 Jan 10;14(1):e0210592. doi: 10.1371/journal.pone.0210592.
Deng et al. (2019). Integrated proteome analyses of wheat glume and awn reveal central drought response proteins under water deficit conditions. J Plant Physiol. 2019 Jan;232:270-283. doi: 10.1016/j.jplph.2018.11.011.
Zhang et al. (2018) DELLA proteins negatively regulate dark-induced senescence and chlorophyll degradation in Arabidopsis through interaction with the transcription factor WRKY6. Plant Cell Rep. 2018 Jul;37(7):981-992. doi: 10.1007/s00299-018-2282-9. Epub 2018 Mar 24. PMID: 29574486.
Wang et al. (2018). A role of GUNs-Involved retrograde signaling in regulating Acetyl-CoA carboxylase 2 in Arabidopsis. Biochem Biophys Res Commun. 2018 Nov 2;505(3):712-719. doi: 10.1016/j.bbrc.2018.09.144.
Borovik and Grabelnych (2018). Mitochondrial alternative cyanide-resistant oxidase is involved in an increase of heat stress tolerance in spring wheat. J Plant Physiol. 2018 Dec;231:310-317. doi: 10.1016/j.jplph.2018.10.007.
Pan et al. (2018). Comparative proteomic investigation of drought responses in foxtail millet. BMC Plant Biol. 2018 Nov 29;18(1):315. doi: 10.1186/s12870-018-1533-9.
Lopez?Calcagno et al. (2018). Overexpressing the H?protein of the glycine cleavage system increases biomass yield in glasshouse and field?grown transgenic tobacco plants. Plant Biotechnology Journal, May 2018.
Li et al. (2018). Comparative proteomic analysis of key proteins during abscisic acid-hydrogen peroxide-induced adventitious rooting in cucumber (Cucumis sativus L.) under drought stress. Journal of Plant Physiology Volume 229, October 2018, Pages 185-194.
Adhikari et al. (2018). Sulfate improves cadmium tolerance by limiting cadmium accumulation, modulation of sulfur metabolism and antioxidant defense system in maize. Environmental and Experimental Botany Volume 153, September 2018, Pages 143-162.
Brandt et al. (2018). Extended darkness induces internal turnover of glucosinolates in Arabidopsis thaliana leaves. PLoS One. 2018 Aug 9;13(8):e0202153. doi: 10.1371/journal.pone.0202153.
Jespersen et al. (2017). Metabolic Effects of Acibenzolar-S-Methyl for Improving Heat or Drought Stress in Creeping Bentgrass. Front Plant Sci. 2017 Jul 11;8:1224. doi: 10.3389/fpls.2017.01224. eCollection 2017. (western blot, Agostis stolonifera cv. ‘Penncross’)
Qiu et al. (2015). Soy 14-3-3 protein SGF14c, a new regulator of tolerance to salt–alkali stress. Plant Biotechnology Reports pp 1-9.
Shaw et al. (2015). Beta-aminobutyric acid mediated drought stress alleviation in maize (Zea mays L.). Environ Sci Pollut Res Int. 2015 Sep 29.
Buxa et al. (2015). Phytoplasma infection in tomato is associated with re-organization of plasma membrane, ER stacks, and actin filaments in sieve elements. ront Plant Sci. 2015; 6: 650. Published online 2015 Aug 19.
Zheng et al. (2014). iTRAQ-based quantitative proteomics analysis revealed alterations of carbohydrate metabolism pathways and mitochondrial proteins in a male sterile cybrid pummelo. J Proteome Res. 2014 May 13.

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