Overview

  • Product name

    HSF1 (pSer326) ELISA kit
    See all HSF1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Buffer 20 11.79ng/ml 2.371%
    Buffer 20 5.54ng/ml 1.019%
    Buffer 20 2.8µg/ml 1.1%
    Inter-assay
    Sample n Mean SD CV%
    Buffer 12.09ng/ml 2.86%
    Buffer 5.65ng/ml 7.08%
    Buffer 2.67ng/ml 2.64%
  • Sample type

    Tissue Extracts, Cell Lysate
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    61 pg/ml
  • Range

    0.39 ng/ml - 12.5 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell Lysate 103 2.15mg/ml - 6mg/ml

  • Assay time

    3h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse, Human
  • Product overview

    Abcam’s HSF1 pSer326 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of HSF1 pSER326 in cell lysates and tissue extracts from Human and mouse origin.


    A HSF1 rat monoclonal antibody has been precoated onto 96-well plates. Standards or test samples are added to the wells, incubated and then washed. A biotinylated HSF1 pSer326 polyclonal antibody is then added, incubated and washed. A Streptavidin-HRP Conjugate is then added and incubated. The plate is washed once more and the TMB substrate is then added which HRP catalyzes, generating a blue coloration after incubation. A stop solution is added which generates conversion to yellow color read at 450 nm which is proportional to the amount of analyte bound.

  • Notes

     

    Compound Cross Reactivity
    HSF2 0.03%
    HSF4 0.05%
  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 100ml
    5X Extraction Reagent 1 x 10ml
    Anti-HSF1 monoclonal Microplate (12 x 8 wells) 1 unit
    Assay Buffer 29 1 x 100ml
    Biotinylated antibody to pSer326 HSF1 1 x 10ml
    Phosphorylated Human HSF1 Standard 2 x 500µl
    Plate Sealer 3 units
    Stop Solution 2 1 x 10ml
    Streptavidin-HRP Conjugate 1 x 10ml
    TMB Substrate 1 x 10ml
  • Research areas

  • Function

    DNA-binding protein that specifically binds heat shock promoter elements (HSE) and activates transcription. In higher eukaryotes, HSF is unable to bind to the HSE unless the cells are heat shocked.
  • Sequence similarities

    Belongs to the HSF family.
  • Domain

    the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
  • Post-translational
    modifications

    Phosphorylated on multiple serine residues, a subset of which are involved in stress-related regulation of transcription activation. Constitutive phosphorylation represses transcriptional activity at normal temperatures. Levels increase on specific residues heat-shock and enhance HSF1 transactivation activity. Phosphorylation on Ser-307 derepresses activation on heat-stress and in combination with Ser-303 phosphorylation appears to be involved in recovery after heat-stress. Phosphorylated on Ser-230 by CAMK2, in vitro. Cadmium also enhances phosphorylation at this site. Phosphorylation on Ser-303 is a prerequisite for HSF1 sumoylation. Phosphorylation on Ser-121 inhibits transactivation and promotes HSP90 binding. Phosphorylation on Thr-142 also mediates transcriptional activity induced by heat. Phosphorylation on Ser-326 plays an important role in heat activation of HSF1 transcriptional activity.
    Sumoylated with SUMO1 and SUMO2 on heat-shock. Heat-inducible sumoylation occurs after 15 min of heat-shock, after which levels decrease and at 4 hours, levels return to control levels. Sumoylation has no effect on HSE binding nor on transcriptional activity. Phosphorylation on Ser-303 is a prerequisite for sumoylation.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasmic during normal growth. On activation, translocates to nuclear stress granules. Colocalizes with SUMO1 in nuclear stress granules.
  • Information by UniProt
  • Alternative names

    • Heat shock factor 1
    • Heat shock factor protein 1
    • Heat shock transcription factor 1
    • HSF 1
    • hsf1
    • HSF1_HUMAN
    • HSTF 1
    • HSTF1
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab136939 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

  • Representative standard curve using ab13639.

  • Parallelism experiments were carried out to determine if the recombinant standard accurately determines [pSer326] HSF1 concentrations in biological matrices. Values were obtained using cell lysates from treated and untreated cells serially diluted in the assay buffer and assessed from a standard curve using four parameter logistic curve fitting.

    The observed values were plotted against the dilution factors. Parallelism of the curves demonstrates that the antigen binding characteristics are similar enough to allow the accurate determination of native analyte levels in diluted samples.

  • HeLa and 3T3 cells were grown to approximately 80% confluency and subjected to heat shock at 42ºC for 2 hours. Extracts were prepared as described in “Sample Preparation” and the levels of HSF1 were determined in the assay. Induction is expressed as the fold change relative to non heat-shocked control samples.

Protocols

References

This product has been referenced in:

See 1 Publication for this product

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