概述

  • 产品名称Anti-Hsp90 alpha抗体
    参阅全部 Hsp90 alpha 一抗
  • 描述
    兔多克隆抗体to Hsp90 alpha
  • 特异性Detects Heat Shock Protein 86 (HSP 86). This antibody does not detect HSP 84.
  • 经测试应用适用于: WB, IP, ICC/IF, IHC-Pmore details
  • 种属���应性
    与反应: Mouse, Rat, Sheep, Human, Non Human Primates
    预测可用于: Rabbit, Horse, Cow, Pig, Chimpanzee, Cynomolgus Monkey, Chinese Hamster
  • 免疫原

    Synthetic peptide corresponding to Mouse Hsp90 alpha aa 2-12 (N terminal).
    Sequence:

    PEETQTQDQPM

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • Primary antibody说明Heat shock proteins (HSP) are expressed in response to various biological stresses, including heat. HSP 90 is a 90 kDa protein that is induced under stress conditions, but is also one of the most abundant cellular proteins found under non-stress conditions. HSP 90 has been found to be associated with a number of other intracellular proteins, including steroid receptors, actin, tubulin, Ah receptor, and some kinases. Studies have shown that murine HSP 90 exists as two forms, HSP 84 and HSP 86, coded by related but separate genes, with 86% homologous amino acid sequences. These forms are analogous to the two forms of human HSP 90, HSP 89 beta and HSP 89 alpha. In an unstressed mouse fibroblast, the basal level of HSP 84 is found to be double that of HSP 86. However, after heat shock, HSP 86 shows a greater increase. Studies also suggest that upon cellular differentiation, the level of HSP 86, but not HSP 84, decreases. HSP 84 and HSP 86, which may be subject to estrogenic regulation, have been found as components of the non-DNA binding form of mouse glucocorticoid receptor, but dissociated from the transformed DNA-binding form.
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab2928 in the following tested applications.

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

应用 Ab评论 说明
WB 1/500 - 1/2000. Detects a band of approximately 86 kDa.
IP Use at an assay dependent concentration.

Use at 2 μg.

Immunoprecipitation experiments with this antibody suggest that HSP90 alpha exists primarily as homodimers in HeLa cells. This antibody is capable of precipitating HSP90 alpha that is complexed with other proteins such as the aryl hydrocarbon (Ah) receptor.

ICC/IF 1/50 - 1/200.
IHC-P Use a concentration of 5 µg/ml.

靶标

Anti-Hsp90 alpha antibody 图像

  • Western blot analysis of HSP90 alpha was performed by loading 50ug of the indicated whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with ab2928 at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG HRP secondary antibody (1:20,000) for at least 1 hour. Chemiluminescent detection was performed.

  • ab2928 labelling Hsp90 alpha in Human colon adenocarcinoma tissue sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Tissues were blocked in 3% BSA in PBST for 30 minutes at room temperature. Tissue sections were incubated with the primary antibody (1:100) for 1 hour. A HRP-conjugated goat anti-rabbit IgG (1:250) was used as the secondary antibody, followed by colorimetric detection using Metal Enhanced DAB Substrate Kit. Tissues were counterstained with hematoxylin and prepped for mouting. Images were taken at 40X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of HSP90 alpha (green) in HeLa cells and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA for 15 minutes at room temperature. Cells were incubated with ab2928 at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488 goat-anti-rabbit IgG secondary antibody (1:400) for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

  • Immunoprecipitation of HSP90 alpha was performed on HeLa cells. Antigen-antibody complexes formed by incubating 500ug whole cell lysate with 2ug of ab2928 overnight on a rocking platform at 4°C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with buffer. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with ab2928 at a dilution of 1:1000 overnight rotating at 4°C. The membrane was washed in TBST, and probed with detection reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) ab2928 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human kidney tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) ab2928 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human breast carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) ab2928 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Western blot of mouse Hsp90 alpha and beta using both ab2928 and ab2927 respectively.

Anti-Hsp90 alpha antibody (ab2928)参考文献

This product has been referenced in:
  • Patel PD  et al. Paralog-selective Hsp90 inhibitors define tumor-specific regulation of HER2. Nat Chem Biol 9:677-84 (2013). Read more (PubMed: 23995768) »
  • Quanz M  et al. Heat shock protein 90a (hsp90a) is phosphorylated in response to DNA damage and accumulates in repair foci. J Biol Chem 287:8803-15 (2012). WB ; Human . Read more (PubMed: 22270370) »

See all 4 Publications for this product

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