HSP70/HSC70 | Heat shock protein 70/Heat shock cognate protein 70 (serum)
AS05 083 | clonality: polyclonal | host: rabbit | reactivity: [global abtibody] for HSP70 and HSC70 in fish, mammals and fungi
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1: 1000 with standard ECL (WB), 1: 10 000 with ECL Advance (GE Healthcare), 1: 1000 (IP)
|Expected | apparent MW||
|Confirmed reactivity||fish, mammals, fungi: Antrodia infirma, A. sinuosa, A. xantha, Catostomus commersonii, Gloeophyllum protractum, G. sepiarium, G. carbonarium, Oligoporus sericiomollis, Phlebia cornea, Junghunia luteoalba|
vertebrates including bovine, hen, mouse, rat; insects including Drosophila melanogaster
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
|Additional information||This antibody is recognizing both, the inducible and the constitutive Hsp70.|
|Selected references||MacLellan et al. (2015). Chaperone roles for TMAO and HSP70 during hyposmotic stress in the spiny dogfish shark (Squalus acanthias). J Comp Physiol B. 2015 Jun 7.
Bessemer et al. (2014). Cardiorespiratory toxicity of environmentally relevant zinc oxide nanoparticles in the freshwater fish Catostomus commersonii. Nanotoxicology. 2014 Nov 27:1-10.
Gorovits et al. (2013). Recruitment of the host plant heat shock protein 70 by tomato yellow leaf curl virus coat protein is required for virus infection. PLoS Once, July 23;8(7).
10 µg of total protein from (1) killi fish muscle, (2) bovine muscle, (3) chicken muscle, (4) rat liver, 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 in 5 % non-fat milk for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5000 (in blocking reagent) 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 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with TMA-6 (Lumigen) 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 5 min.
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