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SampleTypes |
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cell lysates, tissue homogenates
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Specificity |
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recognizes recombinant and native Hsp70, no reactivity with Hsc70, Grp78, E.coli DnaK, M tuberculosis Hsp71 nor with human Hsp60
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Protocol |
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1. Add Hsp70 Standards and samples in duplicate to the pre-coated ready-to-use Hsp70 Immunoassay Plate; incubate then wash.
2. Add Biotin Anti-Hsp70; incubate then wash.
3. Add Avidin-HRP Conjugate; incubate then wash.
4. Develop with TMB Substrate.
5. Add Acid Stop Solution.
6. Measure absorbance at 450nm.
7. Plot Hsp70 standard curve and determine sample concentrations
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Storage |
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at 4°C, except the Recombinant Hsp70 Standard, which should be stored at -20°C.
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References |
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1. Walsh, R.C., Koukoulas, I., et al (2001) Cell Stress Chaperones 6(4): 386-393.
2. Campisi, J., Fleshner, M. (2003) J. Appl. Physiol. 94: 43-52.
3. Campisi, J., Leem, T.H., Greenwood, B.N., et al (2003) Am. J. Physiol. 284: R520-R530
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Notes |
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Inducible Hsp70 in sera was accurately quantified at 5ng/ml in spike recovery studies. Depending on the nature and extent of the experimental
stress, serum levels of inducible Hsp70 may not be within the dynamic range of the assay.
Inducible heat shock protein 70 (Hsp70) is a stress protein whose expression is upregulated when the cell or organism is placed under conditions of stress. Hsp70 is essential for cellular recovery, survival and maintenance of normal cellular function.1 It is also a molecular chaperone that prevents protein aggregation and refolds damaged proteins in response to cellular stress caused by environmental insults, pathogens and disease.
References Citing EKS-700
1. Walsh, R.C., Koukoulas, I., Garnham, A., Moseley, P.L., Hargreaves, M. and Febbraio,
M.A. (2001) Cell Stress Chaperones 6(4): 386-393.
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