| 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 stress or 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. Alternative names: HSP70.1, HSP70-1/HSP70-2, Heat shock 70 kDa protein 8 | | immunogen | | KLH-conjugated synthetic peptide conserved across all known sequences of HSP70 P08107 and HSC70 proteins P11142 | | antibody format | | rabbit; | polyclonal; | | serum | lyophilized; | | | quantity | | 100 µl | - for reconstitution add 100 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), immunoprecipiation (IP) | | related products | | | | additional information | | for detection of plant and algal cytoplasmic hsp70 we recommend following product: AS08 371 | |
application information
|
| recommended dilution | | 1: 1000 with standard ECL (WB), 1: 10 000 with ECL Advance (GE Healthcare), 1: 1000 (IP) |
| expected | apparent MW | | 70 kDa |
| confirmed reactivity | | fish, mammals, fungi: Antrodia infirma, A. sinuosa, A. xantha, Gloeophyllum protractum, G. sepiarium, G. carbonarium, Oligoporus sericiomollis, Phlebia cornea, Junghunia luteoalba |
| predicted reactivity | | 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 | | |
| selected references | | 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). |
application example
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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