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AtpB | Beta subunit of ATP synthase, (chloroplastic + mitochondrial) (rabbit antibody)

AS05 085  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: [global antibody] for plant, green alga, animal and bacterial F-type ATP synthases

AtpB | Beta subunit of ATP synthase, (chloroplastic + mitochondrial) (rabbit antibody) in the group Antibodies for Plant/Algal / Global Antibodies at Agrisera AB (Antibodies for research) (AS05 085)

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product information
Background

ATP synthase is the universal enzyme that synthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient.

Immunogen

KLH-conjugated synthetic peptide derived from available plant, algal (chloroplastic and mitochondrial) and bacterial sequences of beta subunits of F-type ATP synthases, including Arabidopsis thaliana chloroplastic ATP synthase subunit beta UniProt: P19366, TAIR: AtCg00480  and Arabidopsis thaliana mitochondrial ATP synthase subunit beta-1, UniProt: P83483,  TAIR: At5g08670 as well as Chlamydomonas reinhardtii, UniProt: P06541 and A8IQU3

Host Rabbit
Clonality Polyclonal
Clone
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 tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Blue Native-PAGE (BN-PAGE), Immunofluorescence (IF), Western blot (WB)
Related products
AS05 085-10 | anti-AtpB rabbit antibody, smaller pack size of AS05 085
AS05 085PRE | AtpB | beta subunit of ATP synthase, pre-immune serum
AS03 030 | anti-AtpB hen antibody (developed to exactly the same peptide as rabbit antibody)
AS03 030S | ATP synthase subunit beta protein standard for quantitation and positive control
AS08 304 | anti-ATP synthase subunit alpha antibody
AS08 312 | anti-ATP synthase subunit gamma antibody
AS05 071 | anti-ATP synthase subunit c antibody
AS16 3976 | anti-AtpB | Beta subunit of ATP synthase, mitochondrial, rabbit antibody
Additional information

The anti-AtpB antibody will detect the mitochondrial form of the F1 ATP  synthase subcomplex, as well as the chloroplastic CF1 Atp Synthase, and most known bacterial F-type Atp Synthases. Peptide used for antibody production is located in a beta sheet, which is partly exposed near the surface of the AtpB protein.

Anti-AtpB antibody was used as a loading control in Chlamydomonas reinhardtii

application information
Recommended dilution 1 : 100 (IF), 1 : 5000 (BN-PAGE), 1 : 2000-1 : 5 000 (WB)
Expected | apparent MW

53.9 kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)

Confirmed reactivity Arabidopsis thaliana, Bacillus cereus, Bryopsis corticulans, Chlamydomonas reinhardtii, Chlorella vulgaris, Cyanidioschyzon merolae, Escherichia coli, Helicobacter pylori, Hordeum vulgare, Glycine max, Lycopersicum esulentum, Moniliophthora perniciosa, Nannochloropsis salina, Neochloris oleoabundans (chlorophyta), Nicotiana bentamiana, Nicotiana tabacum, Oryza sp. (roots, leafs, pollen), Pheodactylum tricornutum CCAP 1055/1, Plasmodium berghei, Populus sp., Robinia pseudoacacia, Selaginella martensii, Spinacia oleracea, Zea mays

Animal tissues from: cow, chicken, pig, rat, salmon, seal, Locusta migratoria
Predicted reactivity Acinetobacter baumannii, Algae, Cannabis sativa, Clostridium sp., Cyanobacteria, E.coli K-12, Nicotiana plumbaginifolia, Saccharomyces cerevisiae, Salmonella typhimurium, Trichodesmium erythraeum, Triticum aestivum, Vitis vinifera, Zosteria marina, Yrsinia sp.
Not reactive in Archeal V-type ATP synthase
Additional information
Blue Native gel electrophoresis (BN-PAGE) has been performed on samples solubilized with digitonin (4:1) and loaded at 100 µg/well. Gel thickness was 2 mm with 4.5-16 % gradient.

Antibody is recognizing mitochondrial form of AtpB Subota el. al (2011).

This antibody can be used as a loading control for bacteria, Bacillus cereus.
Selected references Atkins and Cross (2018). Inter-Regulation of CDKA/CDK1 and the Plant-Specific Cyclin-Dependent Kinase CDKB in Control of the Chlamydomonas Cell Cycle. Plant Cell. 2018 Jan 24. pii: tpc.00759.2017. doi: 10.1105/tpc.17.00759.
Kanget al. (2017). Increased lipid production by heterologous expression of AtWRI1 transcription factor in Nannochloropsis salina. Biotechnol Biofuels. 2017 Oct 10;10:231. doi: 10.1186/s13068-017-0919-5. Fu et al. (2017). Intracellular spectral recompositioning of light enhances algal photosynthetic efficiency. Sci Adv. 2017 Sep 1;3(9):e1603096. doi: 10.1126/sciadv.1603096. (loading control diatoms)
Shin et al. (2017), Complementation of a mutation in CpSRP43 causing partial truncation of light-harvesting chlorophyll antenna in Chlorella vulgaris. Sci Rep. 2017 Dec 20;7(1):17929. doi10.1038/s41598-017-18221-0.
Wang and Auwerx (2017). Systems Phytohormone Responses to Mitochondrial Proteotoxic Stress. Mol Cell. 2017 Nov 2;68(3):540-551.e5. doi: 10.1016/j.molcel.2017.10.006.
Cantrell and Peers (2017). A mutant of Chlamydomonas without LHCSR maintains high rates of photosynthesis, but has reduced cell division rates in sinusoidal light conditions. PLoS One. 2017 Jun 23;12(6):e0179395. doi: 10.1371/journal.pone.0179395.
Nikolova et al. (2017). Temperature induced remodeling of the photosynthetic machinery tunes photosynthesis in a thermophyllic alga. Plant Physiol. 2017 Mar 7. pii: pp.00110.2017. doi: 10.1104/pp.17.00110.
Jiang et al. (2017). IFT57 stabilizes the assembled intraflagellar transport complex and mediates transport of motility-related flagellar cargo. J Cell Sci. 2017 Mar 1;130(5):879-891. doi: 10.1242/jcs.199117.
Li et al. (2016). Characterization of a novel β-barrel protein (AtOM47) from the mitochondrial outer membrane of Arabidopsis thaliana. J Exp Bot. 2016 Nov;67(21):6061-6075. Epub 2016 Oct 6.
Jallet et al. (2016). Photosynthetic physiology and biomass partitioning in the model diatom Phaeodactylum tricornutum grown in a sinusoidal light regime. Algal research, doi:10.1016/j.algal.2016.05.014.
Ferroni et al. (2016). Light acclimation in the lycophyte Selaginella martensii depends on changes in the amount of photosystems and on the flexibility of the light-harvesting complex II antenna association with both photosystems. New Phytol. 2016 Apr 5. doi: 10.1111/nph.13939.
Heinnickel et al. (2016). Tetratricopeptide repeat protein protects photosystem I from oxidative disruption during assembly. Proc Natl Acad Sci U S A. 2016 Mar 8;113(10):2774-9. doi: 10.1073/pnas.1524040113

Application example

2 µg of total protein extracted with PEB (AS08 300) from  leaf tissue of (1) Arabidopsis thaliana, (2) Spinacia oleracea, (3) Lycopersicon esculentum, (4) Glycine max, (5) Populus sp., (6) Zea mays and (7) Hordeum vulgare were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. In parallel a dilution row (a-g: 10 - 5 - 2.5 - 1.25 - 0.63 - 0.32 - 0.16 µg protein/lane) from sample 1 (Arabidopsis) was processed. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-AtpB (AS08 085, 1:5000, 1h) and secondary anti-rabbit (1:10000, 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 standard ECL (Invitrogen) using a Fuji LAS-3000 CCD (300s, standard sensitivity). 

Application example 2

western blot        detection of AtpB in animal and plant tissue

2 µg of total protein from (1) cow muscle, (2) chicken muscle, (3) pig muscle, (4) rat liver, (5) salmon muscle, (6) seal muscle, (8) Arabidopsis thaliana, (9) Zea mays extracted with Protein Extration Buffer, PEB (AS08 300) and separated on  4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (Agrisera anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according to the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.

M - molecular weight marker


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