HSP70/HSC70 | Heat shock protein 70

Gammarus pulex, Salmo salar (Atlantic salmon), Salvelinus fontinalis (Brook trout)

Application example

 5 µg of total protein from (1) cow muscle, (2) chicken muscle, (3) pig muscle, (4) rat liver, (5) salmon muscle , (6) seal muscle, (7) mummichog heat shock control, (8) mummichog heat shock post-24 hours, extracted with Protein Extration Buffer, PEB (AS08 300), were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) 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 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 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent 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 30 seconds.

Chosen peptide sequence is also conserved in several fish species including: DQ202278.1 HSC70 Fundulus, DQ202279.1 HSP70-1 Fundulus, DQ202280.1 HSP70-2 Fundulus, BT059361.1 HSC70 Salmo salar (atlantic salmon), AB092839.2 HSP70 Carassius auratus (goldfish), BC056709.1 HSP70 Danio rerio (zebrafish) and other animal HSP70 proteins

Heat-shock protein 70 (Hsp70) is the major stress-inducible protein in vertebrates and 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. A killifish is a large family of  various oviparous (egg-laying) cyprinodontiform fish including 1270 different species.