StressXpress®
HSP70 ELISA Kit

Colorimetric detection of HSP70

Catalog No. SKT-105

5 out of 5 based on 1 customer rating
Species Reactivity , Hu , Gt , Dg
Sample Types Cell lysates, Tissue
5 out of 5 based on 1 customer rating
(1 customer review)

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SKU: SKT-105 Categories: , .

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Overview

Product Name HSP70 ELISA Kit
Description

Colorimetric detection of HSP70

Species Reactivity Dog, Human, Goat
Platform Microplate
Sample Types Cell lysates, Tissue
Detection Method Colorimetric Assay
Assay Type Sandwich ELISA (Enzyme-linked Immunosorbent Assay)
Utility ELISA kit used to quantitate HSP70 concentration in samples.
Sensitivity 0.18 ng/ml
Assay Range 0.781 - 50 ng/mL
Incubation Time 30 minutes
Number of Samples 40 samples in duplicate
Other Resources , Kit Booklet , MSDS

Properties

Storage Temperature 4ºC
Shipping Temperature Blue Ice
Product Type ELISA Kits
Assay Overview 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.
Kit Components
Component No. Item Quantity / Size
SKC-105A Anti-Hsp70 Immunoassay Plate 1 Plate
SKC-105B 5X Hsp70 Extraction Reagent 1 vial/10 ml
SKC-105C Recombinant Hsp70 Standard 2 vials
SKC-105D Standard and Sample Diluent 1 vial/ 50 ml
SKC-105E 10X Wash Buffer Concentrate 1 vial/100 ml
SKC-105F Anti-Hsp70 Biotinylated Antibody Concentrate 1 vial/150 µl
SKC-105G Anti-Hsp70 Biotinylated Antibody Diluent 1 vial/ 13 ml
SKC-105H Streptavidin: HRP Concentrate 1 vial/150 µl
SKC-105I Streptavidin: HRP Diluent 1 vial/ 13 ml
SKC-105J TMB Substrate 1 vial/ 13 ml
SKC-105K Stop Solution 1 vial/ 13 ml
SKC-105L Pre-treatment Buffer 1 vial/ 13 ml
Cite This Product HSP70 ELISA Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-105)

Biological Description

Alternative Names HSP70 1 ELISA Kit, HSP70 2 ELISA Kit, HSP70.1 ELISA Kit, HSP72 ELISA Kit, HSPA1 ELISA Kit, HSPA1A ELISA Kit, HSPA1B ELISA Kit
Research Areas Cancer, Heat Shock
Scientific 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 44kDa 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 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 (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. Looking for more information on HSP70? Visit our new HSP70 Scientific Resource Guide at http://www.HSP70.com.
References 1. Zho J. (1998) Cell. 94: 471-480.
2. Boorstein W.R., Ziegelhoffer T. & Craig E.A. (1993) J. Mol. Evol. 38(1): 1-17.
3. Rothman J. (1989) Cell. 59: 591 -601.
4. DeLuca-Flaherty et al. (1990) Cell. 62: 875-887.
5. Bork P., Sander C. & Valencia A. (1992) Proc. Natl Acad. Sci. USA. 89: 7290-7294.
6. Fink A.L. (1999) Physiol. Rev. 79: 425-449.
7. Smith D.F., et al. (1993) Mol. Cell. Biol. 13(2): 869-876.
8. Prapapanich V., et al., (1996) Mol. Cell. Biol. 16(11): 6200-6207.
9. Fernandez-Funez et al., (2000) Nature. 408(6808): 101-106.

Product Images

Typical Standard Curve for the HSP70 ELISA Kit (Enzyme-Linked Immunosorbent Assay) StressXpress® – SKT-105. Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.781 – 50 ng/mL.

Product Citations (4)

Other Citations

Urinary heat shock protein 72 as a biomarker of acute kidney injury in dogs.

Bruchim, Y. et al. (2017) Vet J. [Epub ahead of print].

PubMed ID: N/A Reactivity: Dog

Identification of heat stress-susceptible and -tolerant phenotypes in goats in semiarid tropics.

Rout, P.K., Kaushik, R., Ramachandran, N. and Jindal, S.K. (2017) Animal Production Science. [Epub ahead of print].

PubMed ID: N/A Reactivity: Goat

Differential expression pattern of heat shock protein 70 gene in tissues and heat stress phenotypes in goats during peak heat stress period.

Rout, P.K., Kaushik, R. and Ramachandran, N. (2016) Cell Stress Chaperones. 21(4):645-51.

PubMed ID: 27169748 Reactivity: Goat

Reviews

1 review for HSP70 ELISA Kit

  1. 5 out of 5

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    Based on validation through cited publications.

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