HSP70 | heat shock protein 70 cytoplasmic

325 €

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

PRODUCT INFORMATION IN PDF

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

antibody format

rabbit

polyclonal

serum

lyophilized

quantity

150 µl

for reconstitution add 150 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

western blot (WB), immunoprecipitation (IP)

related products

AS08 348 | anti-chloroplastic HSP70

AS08 347 | anti-mitochondrial HSP70

additional information

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

application information
recommended dilution		1: 3000 with standard ECL on 5 µg of protein per well (WB), 2-3 µl/protein extract of concentration 3-5 mg/ml
expected \| apparent MW		70 kDa
confirmed reactivity		Arabidopsis thaliana, Cucumis sativus, Hordeum vulgare, Silene vulgaris, Solanum lycopersicum, Pinus strobus, Zea mays, algae: Desmodesmus subspicatus, phycobiont: Trebouxia TR1 and TR9
predicted reactivity		dicots including Glycine max, Pisum sativum,Solanum lycopersicum and monocots including Oryza sativa, Triticum aestivum,; trees: Populus balsamifera, moss: Physcomitrella patens, algae including Chlamydomonas reinhardtii and Volvox sp.
not reactive in		no confirmed exceptions from predicted reactivity known in the moment
additional information		not available at the moment
selected references		Alvarez et al. (2012). Different strategies to achieve Pb-tolerance by the two Trebouxia algae coexistin in the lichen Ramalina farinacea. J. Plant Physiol. July 27. Janicka-Russak et al. (2012). Different effect of cadmium and copper on H+-ATPase activity in plasma membrane vesicles from Cucumis sativus roots. J. Exp. Botany, March 2012, ahead of print. Holding (2011). Pyrophosphate dependent fructose-6-phosphate 1-phosphotransferase induction and attenuation of Hsp gene expression during endosperm modification in Quality Protein Maize. Plant Physiol Dec. 8 (ahead of print).

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, Abcam) 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 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, from Sigma ) 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

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