Anti-Hsp90 alpha antibody [2G5.G3] (ab79849)
Key features and details
- Mouse monoclonal [2G5.G3] to Hsp90 alpha
- Suitable for: Flow Cyt, IHC-P, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG1
Overview
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Product name
Anti-Hsp90 alpha antibody [2G5.G3]
See all Hsp90 alpha primary antibodies -
Description
Mouse monoclonal [2G5.G3] to Hsp90 alpha -
Host species
Mouse -
Specificity
ab79849 is Hsp90 alpha specific (>96% alpha specific by ELISA). -
Tested applications
Suitable for: Flow Cyt, IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant full length protein corresponding to Human Hsp90 alpha.
Database link: P07900 -
Positive control
- WB: HEK293T, HAP1 and HeLa whole cell lysate. IHC-P: Human testis tissue. ICC/IF: HaCaT cells. Flow Cyt: HeLa cells.
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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 -20°C. Stable for 12 months at -20°C. -
Storage buffer
Preservative: 0.09% Sodium azide
Constituents: PBS, 50% Glycerol -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
2G5.G3 -
Isotype
IgG1 -
Light chain type
kappa -
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 ab79849 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 |
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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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IHC-P |
Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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WB | (1) |
1/1000 - 1/2000. Predicted molecular weight: 85 kDa.
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Notes |
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Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
IHC-P
Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
WB
1/1000 - 1/2000. Predicted molecular weight: 85 kDa. |
Target
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Function
Molecular chaperone. Has ATPase activity. -
Sequence similarities
Belongs to the heat shock protein 90 family. -
Post-translational
modificationsISGylated. -
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: 3320 Human
- Entrez Gene: 15519 Mouse
- Omim: 140571 Human
- SwissProt: P07900 Human
- SwissProt: P07901 Mouse
- Unigene: 525600 Human
- Unigene: 700831 Human
- Unigene: 1843 Mouse
see all -
Alternative names
- EL52 antibody
- epididymis luminal secretory protein 52 antibody
- Heat shock 86 kDa antibody
see all
Images
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All lanes : Anti-Hsp90 alpha antibody [2G5.G3] (ab79849) 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: 85 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 - ab79849 observed at 90 kDa. Red - loading control ab181602 observed at 36 kDa.
ab79849 Anti-Hsp90 alpha antibody [2G5.G3] was shown to specifically react with Hsp90 alpha 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 alpha knockout samples were subjected to SDS-PAGE. ab79849 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 alpha antibody [2G5.G3] (ab79849)
IHC image of Hsp90 alpha staining in Human Testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab79849, 0.5 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Hsp90 alpha knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)Lanes 1 - 3: Merged signal (red and green). Green - ab79849 observed at 90 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab79849 was shown to specifically react with Hsp90 alpha in wild-type HAP1 cells as signal was lost in Hsp90 alpha knockout cells. Wild-type and Hsp90 alpha knockout samples were subjected to SDS-PAGE. Ab79849 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 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.
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ab79849 at 1/100 dilution staining Hsp90 alpha in human keratinocyte cell line HaCaT by Immunocytochemistry/ immunofluorescence. A Fluorophore conjugated goat anti mouse was used as secondary at 1/50 dilution.
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Overlay histogram showing HeLa cells stained with ab79849 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab79849, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (15)
ab79849 has been referenced in 15 publications.
- Ma J et al. Role of HSP90α in osteoclast formation and osteoporosis development. Exp Ther Med 23:273 (2022). PubMed: 35251339
- Zhao Y et al. Vascular endothelium deploys caveolin-1 to regulate oligodendrogenesis after chronic cerebral ischemia in mice. Nat Commun 13:6813 (2022). PubMed: 36357389
- Antonova A et al. Heat-Shock Protein 90 Controls the Expression of Cell-Cycle Genes by Stabilizing Metazoan-Specific Host-Cell Factor HCFC1. Cell Rep 29:1645-1659.e9 (2019). PubMed: 31693902
- Marrugal Á et al. Impact of Heat Shock Protein 90 Inhibition on the Proteomic Profile of Lung Adenocarcinoma as Measured by Two-Dimensional Electrophoresis Coupled with Mass Spectrometry. Cells 8:N/A (2019). PubMed: 31370342
- Hao J et al. Antiestrogenic Activity of the Xi-Huang Formula for Breast Cancer by Targeting the Estrogen Receptor a. Cell Physiol Biochem 47:2199-2215 (2018). PubMed: 29975948