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HSP70 | Heat shock protein 70 cytoplasmic (100ĩl)

396 €

AS08 371-100 | clonality:polyclonal | host:rabbit | reactivity:A. thaliana, C. sativus, D. subspicatus, E. tef, H. vulgare, P. strobus, S. vulgaris, S. lycopersicum, Trebouxia TR1 and TR9, Z. mays, P. falciparum

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as08 371-100

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product information
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

Immunogen

KLH-conjugated synthetic peptide conserved in known higher plant HSC70 proteins including three isoforms of Arabidopsis thaliana HSC70-1 (NP_ 001119156.1), HSC70-2 (NP_195869.1) and HSC70-3 (NP_187555.1) as well as heat shock inducible Hsp70 of Arabidopsis thaliana AT3g12580/T2E22_110 and At1g16030 and AT3g12580/T2E22_110

Host Rabbit
Clonality Polyclonal
Clone
Purity Serum
Format Lyophilized
Quantity 2 x 50 µl
Reconstitution For reconstitution add 50 µl of sterile water to each tube
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)
Related products

AS08 348 | anti-chloroplastic HSP70
AS08 347 | anti-mitochondrial HSP70

Plant and algal protein extraction buffer

Secondary antibodies

Additional information

Immunoprecipitation protocol using Agrisera anti-Hsp70 cytosolic antibodies can be found here.

application information
Recommended dilution 1 : 3000 5 µg protein/well (WB), 2-3 µl/protein extract of concentration 3-5 mg/ml
Expected | apparent MW

70 kDa

Confirmed reactivity Arabidopsis thaliana, Cucumis sativus, E. teft, Hordeum vulgare, Silene vulgaris, Solanum lycopersicum, Pinus strobus, Zea mays, Desmodesmus subspicatus, phycobiont: Trebouxia TR1 and TR9, Plasmodium falciparum
Predicted reactivity Chlamydomonas reinhardti, Glycine max, Medicago sativa, Nicotiana bethamiana,Oryza sativa, Phaseolus vulgaris, Physcomitrella patens, Pisum sativum, Populus balsamifera, Solanum lycopersicum, Triticum aestivum, Vitis vinifera, Volvox sp.
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references Hazlina et al. (2015). Photoinhibition and Development of Stress Proteins in Macroalgae Irradiated with Ultraviolet Radiation. ASM Sci. J., 7(2), 118–128.
Guggisberg et al. (2014). A sugar phosphatase regulates the methylerythritol phosphate (MEP) pathway in malaria parasites. Nat Commun. 2014 Jul 24;5:4467. doi: 10.1038/ncomms5467.
Liu et al. (2014). Spermidine Enhances Waterlogging Tolerance via Regulation of Antioxidant Defence, Heat Shock Protein Expression and Plasma Membrane H+-ATPase Activity in Zea mays. J. Agronomy and Crop Science, Article first published online: 1 APR 2014, DOI: 10.1111/jac.12058.

Application example

1µg of total protein from (1) Horderum vulgare pre heat shock leaf extracted with PEB (AS08 300), (2) Horderum vulgare post heat shock (2h 40ºC) leaf extracted with PEB (AS08 300), (3) Zea mays pre heat shock total protein leaf extracted with PEB (AS08 300), (4) Zea mays post heat shock (2h 40ºC) total protein leaf extracted with PEB (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 ECL Advance (GE Healthcare)

 

western blot image

 

 

 

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, Hebrew University of Jerusalem, Israel


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