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    products/primary-antibodies/hsp90-antibody-d7a-ab59459.pdf

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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins
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Validated using a knockout cell line

Anti-Hsp90 antibody [D7a] (ab59459)

  • Datasheet
  • SDS
Reviews (9) Submit a question References (16)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)
  • Western blot - Anti-Hsp90 antibody [D7a] (ab59459)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)
  • Western blot - Anti-Hsp90 antibody [D7a] (ab59459)

Key features and details

  • Mouse monoclonal [D7a] to Hsp90
  • Suitable for: WB, IHC-P, IHC-Fr, IP, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Human, Pig
  • Isotype: IgG1

You may also be interested in

Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Protein
Recombinant P. falciparum HSP90 protein (ab222790)
Knockout
Product image
Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (ab266591)

View more associated products

Overview

  • Product name

    Anti-Hsp90 antibody [D7a]
    See all Hsp90 primary antibodies
  • Description

    Mouse monoclonal [D7a] to Hsp90
  • Host species

    Mouse
  • Specificity

    Can isolate complexes of HSP90, Src kinase and cec37

  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IP, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Human, Pig
  • Immunogen

    Full length native protein (purified) corresponding to Chicken Hsp90 alpha aa 1-728. Full length protein HSP90 purified from chicken brain
    Database link: P11501

  • Positive control

    • WB: HEK-293T, HEK-293, HAP1 and HeLa cell lysates. IHC-P: Human colon cancer tissue. Mouse inflamed colon tissue.
  • 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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    D7a
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Heat Shock Proteins
    • Signal Transduction
    • Protein Trafficking
    • Organelle Proteins
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • MMPs
    • Cancer
    • Tumor biomarkers
    • Other
    • Neuroscience
    • Processes

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • KO cell lines

    • Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell line (ab266591)
  • KO cell lysates

    • Human HSP90AA1 (Hsp90 alpha) knockout HEK-293T cell lysate (ab258458)
  • Recombinant Protein

    • Recombinant P. falciparum HSP90 protein (ab222790)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab59459 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 (4)
1/500. Predicted molecular weight: 95 kDa.
IHC-P
1/100000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr (1)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ICC/IF (4)
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Notes
WB
1/500. Predicted molecular weight: 95 kDa.
IHC-P
1/100000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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.
  • 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: 396188 Chicken
    • Entrez Gene: 281832 Cow
    • Entrez Gene: 767874 Cow
    • Entrez Gene: 3320 Human
    • Entrez Gene: 3326 Human
    • Entrez Gene: 15516 Mouse
    • Entrez Gene: 15519 Mouse
    • Entrez Gene: 396742 Pig
    • Entrez Gene: 299331 Rat
    • Entrez Gene: 301252 Rat
    • Omim: 140571 Human
    • Omim: 140572 Human
    • SwissProt: P11501 Chicken
    • SwissProt: Q04619 Chicken
    • SwissProt: Q76LV1 Cow
    • SwissProt: P07900 Human
    • SwissProt: P08238 Human
    • SwissProt: P07901 Mouse
    • SwissProt: P11499 Mouse
    • SwissProt: O02705 Pig
    • SwissProt: P30946 Rabbit
    • SwissProt: P30947 Rabbit
    • SwissProt: P34058 Rat
    • SwissProt: P82995 Rat
    • Unigene: 525600 Human
    • Unigene: 1843 Mouse
    • Unigene: 315997 Mouse
    • Unigene: 341186 Mouse
    • Unigene: 440626 Mouse
    • Unigene: 486334 Mouse
    • Unigene: 119867 Rat
    • Unigene: 232226 Rat
    • Unigene: 3277 Rat
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)

    Immunohistochemistry analysis of ab59459 staining Hsp90 alpha in mouse inflamed colon tissue section at 1/100000 dilution for 12 hours at 4°C using antibody amplifier™ system. Biotin Goat Anti-Mouse antibody was used at 1:2000 for 1 hour at room tempertature. Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at room temperature was used as a counter stain. Localization: Inflammatory cells. Magnification: 40x.

  • Western blot - Anti-Hsp90 antibody [D7a] (ab59459)
    Western blot - Anti-Hsp90 antibody [D7a] (ab59459)
    All lanes : Anti-Hsp90 antibody [D7a] (ab59459) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : HSP90AA1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : HAP1 whole cell lyate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Predicted band size: 95 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab59459 observed at 90 kDa. Red - loading control ab181602 observed at 36 kDa.

     ab59459 Anti-Hsp90 antibody [D7a] was shown to specifically react with Hsp90 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266591 (knockout cell lysate ab258458) was used. Wild-type and Hsp90 knockout samples were subjected to SDS-PAGE. ab59459 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)

    ab59459 staining Hsp90 alpha in human colon cancer tissue section by immunohistochemistry (Bouin's fixed paraffin embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave in citrate buffer. The primary antibody was used at dilution of 1/100,000 dilution using antibody amplifier™ system.

  • Western blot - Anti-Hsp90 antibody [D7a] (ab59459)
    Western blot - Anti-Hsp90 antibody [D7a] (ab59459)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Hsp90 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Hek 293 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab59459 observed at 90 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab59459 was shown to specifically recognize Hsp90 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when Hsp90 knockout cells were examined. Wild-type and Hsp90 knockout samples were subjected to SDS-PAGE. Ab59459 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

Protocols

  • Flow cytometry 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 (16)

Publishing research using ab59459? Please let us know so that we can cite the reference in this datasheet.

ab59459 has been referenced in 16 publications.

  • Snigireva AV  et al. A 2,5-Dihydroxybenzoic Acid-Gelatin Conjugate Inhibits the Basal and Hsp90-Stimulated Migration and Invasion of Tumor Cells. J Funct Biomater 11:N/A (2020). PubMed: 32503118
  • Dhamad AE  et al. 75-kDa glucose-regulated protein (GRP75) is a novel molecular signature for heat stress response in avian species. Am J Physiol Cell Physiol 318:C289-C303 (2020). PubMed: 31800296
  • Güzel C  et al. Quantification of Calcyclin and Heat Shock Protein 90 in Sera from Women with and without Preeclampsia by Mass Spectrometry. Proteomics Clin Appl 13:e1800181 (2019). PubMed: 30417587
  • Zhang S  et al. High expression of HSP90 is associated with poor prognosis in patients with colorectal cancer. PeerJ 7:e7946 (2019). PubMed: 31687275
  • Snigireva AV  et al. Cell surface heparan sulfate proteoglycans are involved in the extracellular Hsp90-stimulated migration and invasion of cancer cells. Cell Stress Chaperones 24:309-322 (2019). PubMed: 30659446
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mice hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Mice hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Oct 20 2021

Western blot abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Hematological cancer cell line lysates)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
Hematological cancer cell line lysates
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 27 2019

Western blot abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mice Hepatocytes)
Gel Running Conditions
Reduced Denaturing (8% SDS-PAGE)
Loading amount
35 µg
Specification
Mice Hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Feb 19 2019

Western blot abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions
Reduced Denaturing (4-15% SDS PAGE)
Loading amount
15 µg
Specification
Rat hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Apr 07 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (DU145)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
DU145
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Mar 31 2017

Western blot abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (DU145)
Gel Running Conditions
Reduced Denaturing (4-15% SDS PAGE)
Loading amount
20 µg
Specification
DU145
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Mar 31 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Hsp90 antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Mar 28 2017

Immunohistochemistry (Frozen sections) abreview for Anti-Hsp90 alpha antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Uterus)
Specification
Uterus
Fixative
Formaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Mukesh Jaiswal

Verified customer

Submitted Apr 12 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-Hsp90 alpha antibody [D7a]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (THP-1 moncyte/macrophage cell line)
Specification
THP-1 moncyte/macrophage cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. James Harris

Verified customer

Submitted Aug 07 2012

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