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Stock availability
In Stock
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Kit s description
Colorimetric detection of HSP70
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Protein target
HSP70
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Scientific context
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 more.
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Primary research area
Cancer, Heat Shock
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Category
Assay ELISA Kits
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Other name
HSP70 1 ELISA Kit, HSP70 2 ELISA Kit, HSP70.1 ELISA Kit, HSP72 ELISA Kit, HSPA1 ELISA Kit, HSPA1A ELISA Kit, HSPA1B ELISA Kit
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Brand name
StressXpress®
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Detection system
Colorimetric Assay
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Assay format
Sandwich ELISA (Enzyme-linked Immunosorbent Assay)
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Main intended usage
ELISA kit used to quantitate HSP70 concentration in samples.
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Brief protocol
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.
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Kit contents
Anti-Hsp70 Immunoassay Plate, 5X Hsp70 Extraction Reagent, Recombinant Hsp70 Standard, Standard and Sample Diluent, 10X Wash Buffer Concentrate, Anti-Hsp70 Biotinylated Antibody Concentrate, Anti-Hsp70 Biotinylated Antibody Diluent, Streptavidin: HRP Concentrate, Streptavidin: HRP Diluent, TMB Substrate, Stop Solution, Pre-treatment Buffer
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Assay precision
Contact us
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Assay cross reactivity
Human
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Samples to be used with this kit
Cell Lysates, Tissue
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Maximum samples to be used with this kit
40 samples in duplicate
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Assay duration
30 minutes
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Kit s sensitivity
0,18 ng/ml
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Assay detection limit
0,781 - 50 ng/ml
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Storage recommendations
4°C
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Shipping recommendations
Blue Ice
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Bibliography
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.
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Release date
1-Nov-2011
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PubMed number
27169748, 25948884
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Tested applications
ELISA
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Tested reactivity
Goat, Human
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Resources available upon request
Kit Booklet, MSDS
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Product image legend
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. Graph of the Typical Standard Curve for the HSP70 ELISA Kit StressXpress - SKT-105
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
500
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Properties
E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays, E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays
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Test
ELISA Enzyme-linked immunosorbent assays Code 90320007 SNOMED
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Gene target
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Gene symbol
HSPA1B, HSPA1A, HSPA14, HSPA1L
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Short name
HSP70 ELISA Kit
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Technique
ELISA kit, ELISA, ELISAs Enzyme-linked immunosorbent assay 90320007 SNOMED code are used by the medical researcher for detection of human, mouse, … proteins are supplied in coated 96 well plates to be stored at +4°C. ELISA test kits can be sandwich ELISA. In sandwich ELISAS Enzyme-linked immunosorbent assay 9we use captor and detector antibodies. In single antibody ELISAs the antigen is coated and only a detector antibody is used. Traditional competition antigen ELISAs are coated with a captor antibody and a competitive antigen is labelled with the chromogen. In this case the ratio on your graph will be inverse proportional with the antigen present in your sample. A sign detected by the Colorimetric detection OD optical density absorbance at 450 nm between 0 and 4 with an ELISA reader for 450 nm absortion. Genprice Inc. the recognized authority tax ID 45-4304622 D-U-N-S number - 078440800. MeSH code E05 478 566 350 170 Enzyme-Linked Immunosorbent Assay. PROMT code 3841327 Immunoabsorbent assay tests. ELISA tests are enzyme linked immunoassays to detect human, mouse or other proteins in serum, plasma, urine or biological fluids. MeSH code E05 478 566 350 170 for Enzyme-Linked Immunosorbent Assay or ELISA kits. Also Enzyme-linked immunoassay, competitive have the SNOMED name 61461007 and the PROMT code for ELISA readers is 3841330. This is the desigation of other diagnostic analyzers and tests.
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Host
antigen assay ELISA kit 1 Kit (96 Wells)
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Alternative name
HSP70 Enzyme-linked immunosorbent assay reagent
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Alternative technique
elisas, kits
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Alternative to gene target
v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog, C-Kit and CD117 and PBT and SCFR, KIT and IDBG-18980 and ENSG00000157404 and 3815, transferase activity, Extracellular, Kit and IDBG-172083 and ENSMUSG00000005672 and 16590, KIT and IDBG-642326 and ENSBTAG00000002699 and 280832
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