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Tag Antibody Set 

HSP70 | Heat shock protein 70 (cytoplasmic)

AS08 371 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. sativus, C. reinhardtii, D. subspicatus, E. tef, G. vermiculophylla, H. vulgare, M. sativa, P. strobus, Salicornia sp., S. italica. S. vulgaris, S. lycopersicum, Trebouxia TR1 and TR9, Z. mays, P. falciparum, V. faba

HSP70 | Heat shock protein 70 (cytoplasmic) in the group Antibodies for Plant/Algal  / Environmental Stress / Heat shock at Agrisera AB (Antibodies for research) (AS08 371)

PRODUCT INFORMATION IN PDF

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Datasheet Product citations Protocols Customer reviews

Product Information

Immunogen
KLH-conjugated synthetic peptide conserved in known higher plant HSC70 proteins including three isoforms of Arabidopsis thaliana HSC70-1 UniProt: F4KCE5 , HSC70-2 UniProt: A0A178UTH3 and HSC70-3 Uniprot: O65719 as well as heat shock inducible Hsp70 of Arabidopsis thaliana TAIR: AT3g12580/T2E22_110 and At1g16030 and AT3g12580/T2E22_110
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 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 Immunoprecipitation (IP), Western blot (WB)
Recommended dilution 1 : 3000 - 1: 10 000, 5 µg protein/well (WB), 2-3 µl/protein extract of concentration 3-5 mg/ml
Expected | apparent MW

70 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Chlamydomonas reinhardtii, Chlamydomonas sp. UWO241, Cucumis sativus, E. teft, Hordeum vulgare, Medicago sativa, Salicornia sp., Silene vulgaris, Solanum lycopersicum, Pinus strobus, Polyscias elegans, Zea mays, algae: Desmodesmus subspicatus, Gracilaria vermiculophylla, phycobiont: Trebouxia TR1 and TR9, Plasmodium falciparum, Setaria italica, Ulva prolifera, Vicia faba
Predicted reactivity Ageratina adenophora, Allium sativum, Arabis alpina, Arachis diogoi, Arundo donax, Brassica napus, brassica rapa subsp. pekinensis, Camellia sinensis, Citrus sinensis, Coffea arabica, Eriobotrya japonica, Gossypium arboretum, Glycine max, Glycine soja, Hordeum vulgare var. distichum, Lotus japonicus, Medicago sativa, medicago truncatula, Musa acuminata subsp. malaccensis, Nicotiana tabacum, Nicotiana bethamiana, Oryza sativa, Phaseolus vulgaris, Physcomitrella patens, Pinus taeda, Pisum sativum, Populus balsamifera, Populus trichocarpa, Salix gilgiana, Saussurea medusa,Solanum tuberosum, Solanum commersonii, Spinacia oleracea, Tragopogon dubius, Tragopogon porrifolius, Triticum aestivum, Vitis vinifera, Volvox sp.
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples

Application example

western blot image


1µg of total protein
from Horderum vulgare pre heat shock leaf (1), Horderum vulgare post heat shock (2h 40ºC) (2), Zea mays pre heat shock total protein leaf (3), Zea mays post heat shock (2h 40ºC) (4), total protein leaf extracted with Agrisera Protein Eextraction Buffer (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF (Milipore). Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-HSP70 antibody (AS08 371, 1:20 000, 1h) and secondary anti-rabbit (1:20 000, 1 h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection reagent with extreme femtogram range.



western blot detection using anti-hsp70 antibody

Protein from Solanum lycopersicum (1) total cell extract ca. 30 -50 µg, (2) and (3) nuclei pellet , (4) and (5) ca. 7 µg of nuclei fraction, (6) and (7) cytoplasmic pellet, (8) ca. 7 µg of cytoplasm fraction, were separated on 10% SDS-PAGE and blotted 1h to nitrocellulose (Schleicher & Schuell). Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-HSP70 antibody (AS08 371, 1:5000, 3h RT). The antibody solution was decanted and the blot was rinsed briefly. Washed 3 times for 15 min in TBS-T at room temperature with agitation. Blot was incubated with a secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 5:000. The blot was washed as above and developed for 1 min with ECL detection reagent according to the manufacturers instructions.

Courtesy Dr Rena Gorovits, The Hebrew University of Jerusalem, Israel
 


 western blot using plant anti-HSP70 antibodies

200 fmoles of HSP70 protein standard product number AS08 371S (1), 1 µg of total protein from samples such as Lycopersicum esculentum leaf (2), Nicotiana tabaccum leaf, (3)Zea mays leaf (4), Hordeum vulgare leaf (5), Arabidopsis thaliana leaf (6) were extracted with Agrisera Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4- 12% Bolt Plus gels,  LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent or 5% non-fat milk dissolved 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: 10 000 (in blocking reagent) for 1h/RT  with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS10 1489, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with chemiluminescence detection reagent in extreme femtogram range,  according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.

Additional information

Contains 0.1% ProClin

This antibody can be used as a marker of cytoplasmic fraction in tomato (Anfoka et al. 2015).

Applied primary antibody dilution in western blot depends upon sensitivity of detection reagents (pico or femtogram for chemiluminescent detection).

Immunoprecipitation protocol using Agrisera anti-Hsp70 cytosolic antibodies, see tab: protocols.

Related products

Related products

AS08 348 | Anti-chloroplastic HSP70, rabbit antibodies
AS08 347 | Anti-mitochondrial HSP70, rabbit antibodies

Plant and algal protein extraction buffer

Secondary antibodies

Background

Background
Heat-shock protein 70 (Hsp70) is the major stress-inducible protein in vertebrates and is highly conserved throughout evolution. It plays a role as a molecular chaperone and is important for allowing cells to cope with acute stressor insult, especially those affecting the protein machinery. Heat shock cognate protein 70 (HSC70), is a highly conserved protein and a member of the family of molecular chaperones.

Product citations

Selected references Lentini et al. (2018). Early responses to cadmium exposure in barley plants: effects on biometric and physiological parameters. Acta Physiologiae Plantarum October 2018, 40:178
Fan et al. (2018). Comparative proteomic analysis of Ulva prolifera response to high temperature stress. Proteome Sci. 2018 Oct 27;16:17. doi: 10.1186/s12953-018-0145-5.
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.
Lentini et al. (2018). Early responses to cadmium exposure in barley plants: effects on biometric and physiological parameters. Acta Physiol Plant (2018) 40: 178. https://doi.org/10.1007/s11738-018-2752-2.
Balážová et al. (2018). Zinc oxide nanoparticles phytotoxicity on halophyte from genus Salicornia. Plant Physiol Biochem. 2018 Sep;130:30-42. doi: 10.1016/j.plaphy.2018.06.013.
Yoon et al. (2018). The subfamily II catalytic subunits of protein phosphatase 2A (PP2A) are involved in cortical microtubule organization. Planta. 2018 Sep 6. doi: 10.1007/s00425-018-3000-0.
Alamri et al. (2018). Nitric oxide-mediated cross-talk of proline and heat shock proteins induce thermotolerance in Vicia faba L. Environmental and Experimental Botany Available online 23 June 2018.
Barghetti et al. (2017). Heat-shock protein 40 is the key farnesylation target in meristem size control, abscisic acid signaling, and drought resistance. Genes Dev. 2017 Nov 15;31(22):2282-2295. doi: 10.1101/gad.301242.117.
Gorovits et al. (2017). The six Tomato yellow leaf curl virus genes expressed individually in tomato induce different levels of plant stress response attenuation. Cell Stress Chaperones. 2017 Mar 21. doi: 10.1007/s12192-017-0766-0.
Fernández-Bautista N. et al. (2017). AtHOP3, a member of the HOP family in Arabidopsis, interacts with BiP and plays a major role in the ER stress response. Plant Cell Environ. 2017 Feb 2. doi: 10.1111/pce.12927.
Hammann et al. (2016). Selection of heat‑shock resistance traits during the invasion of the seaweed Gracilaria vermiculophylla. Marine Biology 163: 104.
Shen et al. (2016). The Arabidopsis polyamine transporter LHR1/PUT3 modulates heat responsive gene expression by enhancing mRNA stability. Plant J. 2016 Aug 19. doi: 10.1111/tpj.13310. [Epub ahead of print]
Gorovits et al. (2016). Tomato yellow leaf curl virus confronts host degradation by sheltering in small/midsized protein aggregates. Virus Res. 2016 Feb 2;213:304-13. doi: 10.1016/j.virusres.2015.11.020. Epub 2015 Dec 1.
Ghandi et al. (2016). Tomato yellow leaf curl virus infection mitigates the heat stress response of plants grown at high temperature. Sci Rep. 2016 Jan 21;6:19715. doi: 10.1038/srep19715.

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