Cancer Biomarker ELISA Assay

HSP70 ELISA Assay Kit

$3,240.00

The HSP70 ELISA Assay Kit is for the detection of human Hsp70 in cell lysates, and tissue extracts. Each kit contains sufficient components to quantitate the Hsp70 concentration in up to 40 samples, tested in duplicate. The HSP70 ELISA Kit is for research use only and should not be used in diagnostic procedures.

SKU: SKT-105 Categories: , ,

HSP70 ELISA Assay Kit

The HSP70 ELISA Assay Kit is For Research Use Only

Sensitivity: 0.18 ng/ml
Dynamic Range: 0.781 – 50 ng/mL
Incubation Time: 30 minutes
Sample Type: Cell lysates, Serum, Tissue, Urine
Sample Size: 50 μL

Product manufactured in Canada by StressMarq.

Assay Background

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 (2). 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 (3). When hsc70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44 kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half (4). The structure of this ATP binding domain displays multiple features of nucleotide binding proteins (5). All hsp70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the hsp70 family recognize and bind to ascent 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 (6). 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.

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Additional Information

Assay Procedure

  1. Prepare Standard and samples in Standard and Sample Diluent.
  2. Add 100 µL of Standard to appropriate wells.
  3. Add 50 µL of Pre-Treatment Buffer to all sample wells.
  4. Add 50 µL of sample to appropriate wells.
  5. Cover plate with Plate Sealer and incubate at 37°C for 2 hours.
  6. Wash plate four times with 1X Wash Buffer.
  7. Add 100 µL of Detection Antibody Working Solution to each well.
  8. Cover plate with Plate Sealer and incubate at 37°C for 2 hours.
  9. Wash plate four times with 1X Wash Buffer as described in step 6.
  10. Add 100 µL of Streptavidin-HRP Working Solution to each well.
  11. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.
  12. Wash plate four times with 1X Wash Buffer as described in step 6.
  13. Add 100 µL of TMB Substrate to each well.
  14. Develop the plate in the dark at room temperature for 30 minutes.
  15. Stop reaction by adding 100 µL of Stop Solution to each well.
  16. Measure absorbance on a plate reader at 450 nm.

Typical Standard Curve

HSP70 ELISA Assay Kit

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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