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    hsp90-alpha-antibody-ab2928.pdf

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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins
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Anti-Hsp90 alpha antibody (ab2928)

  • Datasheet
  • SDS
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Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody (ab2928)
  • Western blot - Anti-Hsp90 alpha antibody (ab2928)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
  • Immunoprecipitation - Anti-Hsp90 alpha antibody (ab2928)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)

Key features and details

  • Rabbit polyclonal to Hsp90 alpha
  • Suitable for: WB, IP, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human, African green monkey
  • Isotype: IgG

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Overview

  • Product name

    Anti-Hsp90 alpha antibody
    See all Hsp90 alpha primary antibodies
  • Description

    Rabbit polyclonal to Hsp90 alpha
  • Host species

    Rabbit
  • Specificity

    Detects Heat Shock Protein 86 (HSP 86). This antibody does not detect HSP 84.
  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    African green monkey
    See all applications and species data
  • Immunogen

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

    PEETQTQDQPM

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: HEK-293T, HeLa, K562, A431, HepG2, COS-7, NIH/3T3, NRK whole cell lysate. ICC HeLa, NIH/3T3, whole cell. IHC-P: Human colon adenocarcinoma, breast carcinoma, tonsil, kidney tissue. IP: HeLa whole cell lyaste.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Primary antibody notes

    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.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Heat Shock Proteins
    • Signal Transduction
    • Protein Trafficking
    • Organelle Proteins
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • MMPs

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Hsp90 alpha protein (Active) (ab80369)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Mouse
Human
IHC-P
Human
IP
Human
WB
Mouse
Rat
Human
African green monkey
Application Abreviews Notes
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.
Notes
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.

Target

  • Function

    Molecular chaperone. Has ATPase activity.
  • Sequence similarities

    Belongs to the heat shock protein 90 family.
  • Post-translational
    modifications

    ISGylated.
  • Cellular localization

    Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Target information above from: UniProt accession P07900 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3320 Human
    • Entrez Gene: 15519 Mouse
    • Entrez Gene: 299331 Rat
    • Omim: 140571 Human
    • SwissProt: P07900 Human
    • SwissProt: P07901 Mouse
    • SwissProt: P82995 Rat
    • Unigene: 525600 Human
    • Unigene: 700831 Human
    • Unigene: 1843 Mouse
    • Unigene: 315997 Mouse
    • Unigene: 341186 Mouse
    • Unigene: 440626 Mouse
    • Unigene: 119867 Rat
    • Unigene: 232226 Rat
    • Unigene: 3277 Rat
    see all
  • Alternative names

    • EL52 antibody
    • epididymis luminal secretory protein 52 antibody
    • Heat shock 86 kDa antibody
    • heat shock 90kD protein 1, alpha antibody
    • Heat shock 90kD protein 1, alpha like 4 antibody
    • heat shock 90kD protein, alpha-like 4 antibody
    • Heat shock 90kDa protein 1 alpha antibody
    • Heat shock protein 90kDa alpha (cytosolic) class A member 1 antibody
    • Heat shock protein HSP 90-alpha antibody
    • HS90A_HUMAN antibody
    • HSP 86 antibody
    • HSP86 antibody
    • Hsp89 antibody
    • HSP89A antibody
    • Hsp90 antibody
    • HSP90A antibody
    • HSP90AA1 antibody
    • HSP90ALPHA antibody
    • HSP90N antibody
    • HSPC1 antibody
    • HSPCA antibody
    • HSPCAL1 antibody
    • HSPCAL4 antibody
    • HSPN antibody
    • LAP 2 antibody
    • LAP2 antibody
    • lipopolysaccharide-associated protein 2 antibody
    • LPS-associated protein 2 antibody
    • Renal carcinoma antigen NY-REN-38 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody (ab2928)
    Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody (ab2928)

    Immunocytochemistry/Immunofluorescence analysis of HSP90 alpha (green) in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and NIH/3T3 (Mouse embryo fibroblast cell line) 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.

  • Western blot - Anti-Hsp90 alpha antibody (ab2928)
    Western blot - Anti-Hsp90 alpha antibody (ab2928)
    All lanes : Anti-Hsp90 alpha antibody (ab2928) at 1/1000 dilution

    Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate
    Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
    Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 6 : COS-7 (African green monkey kidney fibroblast-like cell line) whole cell lysate
    Lane 7 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
    Lane 8 : NRK (Rat kidney normal tissue) whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit IgG HRP secondary antibody at 1/20000 dilution


    Western blot analysis of HSP90 alpha was performed by loading samples 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 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a secondary antibody for at least 1 hour. Chemiluminescent detection was performed.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)

    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.

  • Immunoprecipitation - Anti-Hsp90 alpha antibody (ab2928)
    Immunoprecipitation - Anti-Hsp90 alpha antibody (ab2928)

    Immunoprecipitation of HSP90 alpha was performed on HeLa (Human epithelial cell line from cervix adenocarcinoma) 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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)

    Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human tonsil tissue. 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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)

    Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human kidney tissue. 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.

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (19)

Publishing research using ab2928? Please let us know so that we can cite the reference in this datasheet.

ab2928 has been referenced in 19 publications.

  • Zhang XH  et al. Heat shock protein 90 relieves heat stress damage of myocardial cells by regulating Akt and PKM2 signaling in vivo. Int J Mol Med 45:1888-1908 (2020). PubMed: 32236591
  • Inda MC  et al. The epichaperome is a mediator of toxic hippocampal stress and leads to protein connectivity-based dysfunction. Nat Commun 11:319 (2020). PubMed: 31949159
  • Gorska-Ponikowska M  et al. The Major Heat Shock Proteins, Hsp70 and Hsp90, in 2-Methoxyestradiol-Mediated Osteosarcoma Cell Death Model. Int J Mol Sci 21:N/A (2020). PubMed: 31963524
  • Lin C  et al. Fever Promotes T Lymphocyte Trafficking via a Thermal Sensory Pathway Involving Heat Shock Protein 90 and a4 Integrins. Immunity 50:137-151.e6 (2019). PubMed: 30650373
  • Pan C  et al. Hsp90B enhances MAST1-mediated cisplatin resistance by protecting MAST1 from proteosomal degradation. J Clin Invest 129:4110-4123 (2019). PubMed: 31449053
View all Publications for this product

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