Overview

  • Product name

    Anti-HSF1 (phospho S326) antibody [EP1713Y]
    See all HSF1 primary antibodies
  • Description

    Rabbit monoclonal [EP1713Y] to HSF1 (phospho S326)
  • Host species

    Rabbit
  • Specificity

    ab76076 detects HSF1 phosphorylated on Ser326.
  • Tested applications

    Suitable for: Dot blot, WBmore details
    Unsuitable for: Flow Cyt,ICC,IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    corresponding to Human HSF1 aa 300-400.

  • Positive control

    • WB: HeLa cell lysates and HeLa cell lysate treated with heat (44ºC)
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab76076 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
Dot blot Use a concentration of 0.338 µg/ml.
WB 1/5000 - 1/10000. Predicted molecular weight: 57 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,ICC,IHC-P or IP.
  • Target

    • 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
    • Database links

    • Alternative names

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

    Images

    • All lanes : Anti-HSF1 (phospho S326) antibody [EP1713Y] (ab76076) at 0.169 µg/ml

      Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HeLa treated with heat at 43? for 30 minutes whole cell lysate
      Lane 3 : HeLa treated with heat at 43? for 30 minutes whole cell lysate. Then the membrane was incubated with alkaline phosphatase.

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.02 µg/ml

      Predicted band size: 57 kDa
      Observed band size: 82 kDa
      why is the actual band size different from the predicted?



      Blocking/Diluting Buffer and concentration: 5% NFDM /TBST

    • All lanes : Anti-HSF1 (phospho S326) antibody [EP1713Y] (ab76076) at 1/10000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with heat (42 ?) whole cell lysates
      Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with heat (42 ?) whole cell lysates. Then the membrane was incubated with phosphatase

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 57 kDa
      Observed band size: 82 kDa why is the actual band size different from the predicted?

    • Dot blot analysis of HSF1 (phospho S326) peptide (Lane 1), HSF1 non-phospho peptide (Lane 2) labelling HSF1 (phospho S326) with purified ab76076 at 0.338 ug/mL. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at 0.01 ug/mL.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-HSF1 (phospho S326) antibody [EP1713Y] (ab76076) at 1/10000 dilution (unpurified)

      Lane 1 : HeLa cell lysates (untreated)
      Lane 2 : HeLa cell lysates, treated with heat (44o C)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP at 1/1000 dilution

      Predicted band size: 57 kDa
      Observed band size: 82 kDa why is the actual band size different from the predicted?

    References

    This product has been referenced in:

    • Girard PM  et al. Differential correlations between changes to glutathione redox state, protein ubiquitination, and stress-inducible HSPA chaperone expression after different types of oxidative stress. Cell Stress Chaperones N/A:N/A (2018). Read more (PubMed: 29754332) »
    • Li L  et al. GKAP Acts as a Genetic Modulator of NMDAR Signaling to Govern Invasive Tumor Growth. Cancer Cell 33:736-751.e5 (2018). Read more (PubMed: 29606348) »
    See all 25 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Answer

    Thank you for contacting us and bringing this to our attention.


    We have tested both products (ab76076 and ab81281) in mouse and rat samples, via WB.
    The samples we tested were brain, heart, kidney and spleen for each, in mouse and rat. We only saw a few non-specific bands (with both products) in mouse kidney. No bands were observed in the other lysates.
    We do not recommend these products for use in mouse and rat based on this testing.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (colon)
    Loading amount
    20 µg
    Specification
    colon
    Treatment
    heat shock
    Gel Running Conditions
    Reduced Denaturing (7.5%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

    Ms. Young Ju Yoon

    Verified customer

    Submitted Sep 04 2009

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