Recombinant
RabMAb

Recombinant Anti-Hsp90 antibody [EPR16621-67] (ab203126)

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Overview

  • Product name

    Anti-Hsp90 antibody [EPR16621-67]
    See all Hsp90 primary antibodies

  • Description

    Rabbit monoclonal [EPR16621-67] to Hsp90

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Hsp90 aa 500 to the C-terminus. The exact sequence is proprietary. Also UniProt ID: P07900
    Database link: P08238

  • Positive control

    • WB: Hsp90 alpha recombinant protein fragment (GST-tag): aa533-732; Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724 ; Human fetal brain, fetal heart and fetal kidney lysates; Mouse and rat brain, heart and kidney lysates; HeLa, HEK-293, K562, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Human testis, Human lung cancer, mouse cerebral cortex and rat testis tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow cytometry: Jurkat cells. IP: HeLa whole cell lysate.

  • General notes

     

     

     

     

    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.

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 long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR16621-67

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab203126 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
ICC/IF 1/200.

Methanol fixed cells.
IP 1/100.
Flow Cyt 1/350.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 85, 83 kDa).

Target

  • Function

    Molecular chaperone that promotes the maturation, structural maintenance and proper regulation of specific target proteins involved for instance in cell cycle control and signal transduction. Undergoes a functional cycle that is linked to its ATPase activity. This cycle probably induces conformational changes in the client proteins, thereby causing their activation. Interacts dynamically with various co-chaperones that modulate its substrate recognition, ATPase cycle and chaperone function.

  • Sequence similarities

    Belongs to the heat shock protein 90 family.

  • Domain

    The TPR repeat-binding motif mediates interaction with TPR repeat-containing proteins like the co-chaperone STUB1.

  • Post-translational
    modifications

    ISGylated.
    S-nitrosylated; negatively regulates the ATPase activity and the activation of eNOS by HSP90AA1.

  • Cellular localization

    Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Heat shock 86 kDa antibody
    • Heat shock protein 90kDa alpha cytosolic class A member 1 antibody
    • Heat shock protein 90kDa alpha cytosolic class B member 1 antibody
    • Heat shock protein HSP 90 alpha antibody
    • Heat shock protein HSP 90 beta antibody
    • Heat shock protein HSP 90-alpha antibody
    • HS90A_HUMAN antibody
    • HSP 84 antibody
    • HSP 86 antibody
    • Hsp 90 antibody
    • HSP86 antibody
    • HSP90A antibody
    • HSP90AA1 antibody
    • HSP90AB1 antibody
    • HSP90B antibody
    • HSPC1 antibody
    • HSPC2 antibody
    • HSPCAL1 antibody
    • HSPCAL4 antibody
    • Renal carcinoma antigen NY-REN-38 antibody
    see all

Images

  • Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 85, 83 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Immunocytochemistry/Immunofluorescence analysis of 100% methanol-fixed NIH/3T3 (Mouse embryonic fibroblast) cells labeling Hsp90 alpha + beta with ab203126 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Control - PBS instead of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on germ cells of Human testis is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Immunocytochemistry/Immunofluorescence analysis of 100% methanol-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Hsp90 alpha + beta with ab203126 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Control - PBS instead of primary antibody.

  • Flow Cytometry - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Flow Cytometry - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Hsp90 alpha + beta with ab203126 at 1/350 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunoprecipitation - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Hsp90 alpha + beta was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab203126 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab203126 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate 10 µg (Input).

    Lane 2: ab203126 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203126 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on tumor cells of Human lung cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

    Lane 1 : Hsp90 alpha recombinant protein fragment (GST-tag): aa533-732
    Lane 2 : Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size: 85, 83 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Hsp90 alpha and Hsp90 beta recombinant protein fragments were made in house.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and nucleus staining on neuron of mouse cerebral cortex is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 85, 83 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

    Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on germ cells of rat testis is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)
    Lanes 1 & 3-4 : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/100000 dilution
    Lane 2 : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size: 85, 83 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Liu R  et al. Inhibition of lncRNA NEAT1 suppresses the inflammatory response in IBD by modulating the intestinal epithelial barrier and by exosome-mediated polarization of macrophages. Int J Mol Med 42:2903-2913 (2018). Read more (PubMed: 30132508) »
  • Liu X  et al. Effects of ginsenoside Rb1 on oxidative stress injury in rat spinal cords by regulating the eNOS/Nrf2/HO-1 signaling pathway. Exp Ther Med 16:1079-1086 (2018). Read more (PubMed: 30116359) »
See all 3 Publications for this product

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