Overview

  • Product name
    Human HSP70 ELISA Kit
    See all HSP70 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.6%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 6.1%
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    2.5 U/ml
  • Range
    7.81 U/ml - 500 U/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 90.8 79.7% - 96.6%
    Tissue Culture Media 69.5 67.6% - 70.5%
    Goat Serum 67.9 67% - 68.6%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    HSP70 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of HSP70 protein in human cell and tissue extract samples.


    The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    HSP70 belongs to the heat shock protein 70 family. In cooperation with other chaperones, HSP70s stabilize preexistent proteins against aggregation and mediate the folding of newly translated polypeptides in the cytosol as well as within organelles. These chaperones participate in all these processes through their ability to recognize nonnative conformations of other proteins. They bind extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In case of rotavirus A infection, HSP70 serves as a post-attachment receptor for the virus to facilitate entry into the cell. HSP70 is induced by heat shock.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab187399 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD, n=2) are graphed.

  • Background-subtracted data values (mean +/- SD, n=2) are graphed.

  • Cell extracts (250 µg/mL) prepared from un-induced (control) and heat-shocked (HS, 30 min at 42ºC, followed by recovery period, as indicated, at 37ºC) cells were analyzed with this kit. The concentrations of HSP70 were interpolated from data values using HSP70 standard curve and graphed in units (U) of HSP70 per mg of extract (mean, n=2).

  • Cell extracts (40 µg) prepared from un-induced (lane 3, Jurkat; lane 7, HeLa) cells or 30 min at 42ºC heat-shocked, then recovered at 37ºC for variable time periods (lane 4, 1 hour; lane 5, 4 hours; lane 6, 6.5 hours) Jurkat cells were analyzed by Western blotting using the capture antibody of this kit. The Human recombinant HSP70 protein standard was analyzed as well (lane 2). Note that this antibody detects a single band of 70 kDa and that the HSP70 levels obtained by this analysis correlate with the result obtained with the use of this kit.

  • Cell extracts (40 µg) prepared from un-induced (lane 3, Jurkat; lane 7, HeLa) cells or 30 min at 42ºC heat-shocked, then recovered at 37ºC for variable time periods (lane 4, 1 hour; lane 5, 4 hours; lane 6, 6.5 hours) Jurkat cells were analyzed by Western blotting using the detector antibody of this kit. The Human recombinant HSP70 protein standard was analyzed as well (lane 2). Note that this antibody detects a single band of 70 kDa and that the HSP70 levels obtained by this analysis correlate with the result obtained with the use of this kit.

Protocols

References

ab187399 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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