Hsp70 (Heat Shock Protein 70) BioAssay™ ELISA Kit (High Sensitivity)
Product Sizes
96Tests
£1023.00
052349-96TESTS
About this Product
- SKU:
- 052349
- Extra Details:
- This Heat Shock Protein 70 (Hsp70) BioAssay™ ELISA Kit is a colorimetric sandwich immunoassay for the quantitative determination of HSP70 in canine, human, monkey, mouse and rat cell lysates, plasma, serum and tissue. HSP70 genes encode abundant heat-inducible 70-kDa HSPs (HSP70s). In most eukaryotes HSP70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 5O% identity. The N-terminal two thirds of HSP70s are more conserved than the C-terminal third. HSP70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides. When HSC70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half. The structure of this ATP binding domain displays multiple features of nucleotide binding proteins. All HSP70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the HSP70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein. The universal ability of HSP70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport. Sensitivity: 0.038ng/ml Detection Range: 0.42-27ng/ml Assay Procedure: 1. Prepare Standard and samples in Standard and Sample Diluent. 2. Add 100 µL of Standard or sample to appropriate wells. 3. Cover plate with Plate Sealer and incubate at 37°C for 1 hour with shaking at 600rpm. 4. Wash plate four times with 1X Wash Buffer. 5. Add 100 µL of Biotinylated Antibody Working Solution to each well. 6. Cover plate with Plate Sealer and incubate at at 37°C for 1 hour with shaking at 600rpm. 7. Wash plate four times with 1X Wash Buffer. 8. Add 100 µL of Streptavidin-HRP Working Solution to each well. 9. Cover plate with Plate Sealer and incubate at 37°C for 30 minutes with shaking at 600rpm. 10. Wash plate four times with 1X Wash Buffer. 11. Add 100 µL of TMB Substrate to each well. 12. Develop the plate in the dark at room temperature for 30 minutes. 13. Stop reaction by adding 100 µL of Stop Solution to each well. 14. Measure absorbance on a plate reader at 450 nm. Kit Components: Microtiter Strip 1x96 wells, Pre-coated with Hsp70 antibody. Recombinant Hsp70 Standard, 2xvial Standard and Sample Diluent, 1x50ml Wash Buffer, 10X, 1x100ml. Hsp70 antibody Concentrate (Biotin), 1x150ul Hsp70 antibody Diluent (Biotin), 1x150ul Streptavidin: HRP Concentrate, 1x50ul Streptavidin: HRP Diluent, 1x13ml TMB Substrate: 1x13ml Stop Solution: 1x13ml Storage and Stability: Standard and Streptavidin HRP Concentrate should be stored at -20°C. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
- Shipping Conditions:
- Blue Ice
- Storage Conditions:
- 4[o]C/-20[o]C
- Type:
- Assay Kits:Assays
