Overview

  • Product name

    Anti-Hsp90 beta antibody [EPR16621]
    See all Hsp90 beta primary antibodies
  • Description

    Rabbit monoclonal [EPR16621] to Hsp90 beta
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human Hsp90 beta aa 500 to the C-terminus.
    Database link: P08238

  • Positive control

    • WB: Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724; HeLa, 293T, K562, Jurkat whole cell lysates. Mouse brain, heart, kidney, spleen. Rat brain, heart, kidney, spleen. C6, RAW 264.7, PC-12, NIH/3T3 whole cell lysates. IHC-P: Human, Mouse, Rat cerebral cortex tissue; Human testis tissue. ICC/IF: HeLa cells. IP: HeLa cells. Flow: HeLa cells.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

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: 0.05% BSA, 40% Glycerol, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR16621
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab203085 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
WB 1/5000. Detects a band of approximately 90 kDa (predicted molecular weight: 85 kDa).
ICC/IF 1/250.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/70.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Post-translational
    modifications

    Ubiquitinated in the presence of STUB1-UBE2D1 complex (in vitro).
    ISGylated.
    S-nitrosylated; negatively regulates the ATPase activity.
  • Cellular localization

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

  • Alternative names

    • 90 kda heat shock protein beta HSP90 beta antibody
    • D6S182 antibody
    • FLJ26984 antibody
    • Heat shock 84 kDa antibody
    • Heat shock 90kD protein 1, beta antibody
    • Heat shock 90kDa protein 1 beta antibody
    • Heat shock protein 90 alpha family class B member 1 antibody
    • Heat shock protein 90 kDa antibody
    • Heat shock protein 90kDa alpha (cytosolic) class B member 1 antibody
    • Heat shock protein 90kDa alpha family class B member 1 antibody
    • Heat shock protein beta antibody
    • Heat shock protein HSP 90 beta antibody
    • Heat shock protein HSP 90-beta antibody
    • HS90B_HUMAN antibody
    • HSP 84 antibody
    • HSP 90 antibody
    • HSP 90 b antibody
    • HSP 90b antibody
    • HSP84 antibody
    • HSP90 BETA antibody
    • hsp90ab1 antibody
    • HSP90B antibody
    • HSPC2 antibody
    • HSPCB antibody
    see all

Images

  • All lanes : Anti-Hsp90 beta antibody [EPR16621] (ab203085) 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 (Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated)

    Predicted band size: 85 kDa


    Exposure time: 3 minutes


    Blocking and Diluting buffer and concentration: 5%
    NFDM/TBST.

    ab203085 only recognizes Hsp90 beta based on recombinant protein WB result.

  • Immunofluorescence analysis of 4% paraformaldehyde fixed HeLa cells (Human epithelial cells from cervix adenocarcinoma) labeling Hsp90 beta with ab203085 at a 1/250 dilution showing cytoplasmic staining. Permeabilisation using 0.1% tritonX-100.

    Secondary ab: Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/200 dilution. Counter stain is labeling tubulin (red) with ab7291 at 1/500 dilution with secondary antibody Anti-Mouse AlexaFluor® 594 (ab150120) at 1/400 dilution. DAPI stains the nucleus in blue. -ve control 1 is ab178854 at 1/250 dilution, Anti-Mouse AlexaFluor® 594 (ab150120) at 1/400 dilution. -ve control 2 is ab7291 at 1/500 dilution, Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/200 dilution.

  • All lanes : Anti-Hsp90 beta antibody [EPR16621] (ab203085) at 1/20000 dilution

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

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

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



    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM /TBST

  • All lanes : Anti-Hsp90 beta antibody [EPR16621] (ab203085) at 1/5000 dilution

    Lane 1 : Mouse brain
    Lane 2 : Mouse heart
    Lane 3 : Mouse kidney
    Lane 4 : Mouse spleen
    Lane 5 : Rat brain
    Lane 6 : Rat heart
    Lane 7 : Rat kidney
    Lane 8 : Rat spleen
    Lane 9 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 10 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 12 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

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



    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM /TBST

  • Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Hsp90 beta with ab203085 at 1/500 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin. Negative control also shown using PBS instead of primary antibody and secondary as above.

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

  • Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Hsp90 beta with ab203085 at 1/500 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on neuron of cerebral cortex is observed. This protein is synthesized in cytoplasm, but it can shuttle between cytoplasm and nucleus depend on the cell status. Counter stained with hematoxylin. Negative control also shown using PBS instead of primary antibody and secondary as above.

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

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling Hsp90 beta with ab203085 at 1/500 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on neuron of cerebral cortex is observed. This protein is synthesized in cytoplasm, but it can shuttle between cytoplasm and nucleus depending on the cell status. Counter stained with hematoxylin. Negative control also shown using PBS instead of primary antibody and secondary as above.

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

  • Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling Hsp90 beta with ab203085 at 1/500 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on neuron of cerebral cortex is observed. This protein is synthesized in cytoplasm, but it can shuttle between cytoplasm and nucleus depending on the cell status. Counter stained with hematoxylin. Negative control also shown using PBS instead of primary antibody and secondary as above.

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

  • Hsp90 beta was immunoprecipitated from 1mg of Hela (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab203085 at 1/70 dilution. Western blot was performed of the immunoprecipitate using ab203085 at 1/5000 dilution. Goat Anti-Rabbit IgG, (H+L), peroxidase conjugated, was used as secondary antibody at 1:1000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM/TBST.

  • Flow cytometry analysis of 2% paraformaldehyde fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells using ab203085 at a 1/100 dilution (red) or a Rabbit monoclonal IgG (negative) (black). Goat anti rabbit IgG (FITC) secondary used at a 1/150 dilution.

References

This product has been referenced in:

  • Wang H  et al. Hsp90ab1 stabilizes LRP5 to promote epithelial-mesenchymal transition via activating of AKT and Wnt/ß-catenin signaling pathways in gastric cancer progression. Oncogene 38:1489-1507 (2019). Read more (PubMed: 30305727) »
See 1 Publication for this product

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