Anti-Hsp90 antibody [D7a] (ab59459)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)](https://www.abcam.com/ps/products/59/ab59459/Images/ab59459-450901-AntiHsp90-antibody-IHC.png "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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
Overview
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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
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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.
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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
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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
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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.
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| IHC-P |
1/100000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
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| IHC-Fr | (1) |
Use at an assay dependent concentration.
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| IP |
Use at an assay dependent concentration.
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| ICC/IF | (4) |
Use at an assay dependent concentration.
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| 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. |
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IHC-Fr
Use at an assay dependent concentration. |
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IP
Use at an assay dependent concentration. |
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ICC/IF
Use at an assay dependent concentration. |
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Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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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
modificationsISGylated.
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
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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
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
see all
Images
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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.
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![Western blot - Anti-Hsp90 antibody [D7a] (ab59459)](https://www.abcam.com/ps/products/59/ab59459/Images/ab59459-412757-anti-hsp90-antibody-d7a-western-blot-human-knockout-lysate.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [D7a] (ab59459)](https://www.abcam.com/ps/products/59/ab59459/Images/Hsp90-alpha-Primary-antibodies-ab59459-1.jpg)
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.
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![Western blot - Anti-Hsp90 antibody [D7a] (ab59459)](https://www.abcam.com/ps/products/59/ab59459/Images/ab59459-286271-ab59459KOWb.jpg)
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
Datasheets and documents
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SDS download
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Datasheet download
References (16)
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