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重组RabMAb

重组Anti-HSF1 (phospho S303 + S307)抗体[EP1711Y] (ab81281)

Submit a review Q&A (1)References (5)
Western blot - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
  • Western blot - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
  • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
  • Flow Cytometry (Intracellular) - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1711Y] to HSF1 (phospho S303 + S307)
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-HSF1 (phospho S303 + S307)抗体[EP1711Y]
    参阅全部 HSF1 一抗

  • 描述

    兔单克隆抗体[EP1711Y] to HSF1 (phospho S303 + S307)

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
    不适用于: Flow Cyt or IP

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab78795)

  • 阳性对照

    • WB: HeLa and K-562 whole cell lysates. IHC-P: Human breast carcinoma. ICC/IF: HeLa cells. Flow Cyt (Intra): HeLa cells.

  • 常规说明

    This product has switched from a hybridoma to recombinant production method on 30th May 2023.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab81281于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/50000. Predicted molecular weight: 57 kDa.Can be blocked with Recombinant Human HSF1 protein (ab78795).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
Flow Cyt (Intra)
1/90.
说明
WB
1/50000. Predicted molecular weight: 57 kDa.Can be blocked with Recombinant Human HSF1 protein (ab78795).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
Flow Cyt (Intra)
1/90.
应用说明
Is unsuitable for Flow Cyt or IP.

靶标

  • 功能

    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.

  • 序列相似性

    Belongs to the HSF family.

  • 结构域

    the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.

  • 翻译后修饰

    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.

  • 细胞定位

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

  • 数据库链接

    • 别名

      • 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

    图片

    • Western blot - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      Western blot - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      All lanes : Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281) at 1/50000 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
      Lane 2 : HeLa whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour

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

      Predicted band size: 57 kDa
      Observed band size: 76 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      Western blot - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      All lanes : Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281) at 1/50000 dilution

      Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg
      Lane 2 : K-562 whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour

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

      Predicted band size: 57 kDa
      Observed band size: 76 kDa why is the actual band size different from the predicted?


      Exposure time: 100 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)

      Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling HSF1 with ab81281 at 1/2000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Human breast carcinoma without alkaline phosphatase treatment (A) compared to no signal detected when treated with alkaline phosphatase (B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval Solution2) for 30 mins. Counterstained with hematoxylin.

    • Immunocytochemistry/ Immunofluorescence - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      Immunocytochemistry/ Immunofluorescence - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling HSF1 with ab81281 at 1/1000 dilution. Confocal image showing cytoplasmic and membranous staining in subsets of HeLa cells. ab195889 AlexaFluor® 594-conjugated Anti-alpha Tubulin was used to counterstain tubulin at 1/200 dilution. The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)
      Flow Cytometry (Intracellular) - Anti-HSF1 (phospho S303 + S307) antibody [EP1711Y] (ab81281)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa cells labelling HSF1 with ab81281 at 1/90 dilution (Red) compared with an isotype control (Black) and an unlabelled control (Cell without incubation with primary antibody and secondary antibody (Blue)). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody.

    文献 (5)

    发表研究结果有使用 ab81281？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab81281 被引用在 5 文献中.

    • Mun GI  et al. Decreased expression of FBXW7 by ERK1/2 activation in drug-resistant cancer cells confers transcriptional activation of MDR1 by suppression of ubiquitin degradation of HSF1. Cell Death Dis 11:395 (2020). PubMed: 32457290
    • Hwang HV  et al. Impaired proteostasis in senescent vascular endothelial cells: a perspective on estrogen and oxidative stress in the aging vasculature. Am J Physiol Heart Circ Physiol 316:H421-H429 (2019). PubMed: 30499713
    • Zhao J  et al. Association between integrin-linked kinase and hyperthermia in oral squamous cell carcinoma. Oncol Lett 14:7705-7714 (2017). PubMed: 29250172
    • Asano Y  et al. IER5 generates a novel hypo-phosphorylated active form of HSF1 and contributes to tumorigenesis. Sci Rep 6:19174 (2016). WB . PubMed: 26754925
    • Zhao J  et al. Correlation between the expression of Id-1 and hyperthermia-associated molecules in oral squamous cell carcinoma. J Clin Pathol 66:758-63 (2013). IHC ; Human . PubMed: 23723304

    客户评价及客户问答

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    Question

    Inquiry: this antibody, as written on the datasheet, does not react with mouse tissue and murine species.... ONLY human (also on epitomics website) Lindquist (Cell and JCI...) has published the usage of this ab in mice cell line and tissues..... please could you confirm that ab76076 and ab81281 work on WB on mice cells and mice tissues, thank you best regards

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    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.

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