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product information
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| background |
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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. |
| immunogen |
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KLH-conjugated C-terminal synthetic peptide conserved in hsc/hsp70 sequences from a wide range of animal species |
| antibody format |
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rabbit |
polyclonal |
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serum, |
lyophilized |
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| quantity |
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200 µl, |
for reconstitution add 200 µl, of sterile water. |
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| storage |
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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 |
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western blot (WB) |
| related products |
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collection of antibodies to fish related targets |
| additional information |
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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 |
application information
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| recommended dilution |
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1:10 000 with ECL Advance (WB) |
| expected | apparent MW |
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70 kDa |
| confirmed reactivity |
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salmon (Salmo salar) , rainbow trout (Oncorhynchus mykiss), cow, chicken, pig, rat, seal, mummichog |
| predicted reactivity |
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atlantic salmon (Salmo salar), brook trout (Salvelinus fontinalis) |
| not reactive in |
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no confirmed exceptions from predicted reactivity known in the moment |
| additional information |
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not available at the moment |
| selected references |
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Chandra et al. (2012). Sustained high temperature increases the vitellogenin response to 17 alpha-ethynylestradiol in mummichog (Fundulus
heteroclitus). Aquatic toxicology. |
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.
||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at support@agrisera.com
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