Product nameAnti-Hsp90 alpha antibody [2G5.G3]
See all Hsp90 alpha primary antibodies
DescriptionMouse monoclonal [2G5.G3] to Hsp90 alpha
Specificityab79849 is Hsp90 alpha specific (>96% alpha specific by ELISA).
Tested applicationsSuitable for: Flow Cyt, IHC-Fr, IHC-P, ELISA, WB, IP, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Human Hsp90 alpha
- WB: HAP1 and HeLa whole cell lysate. IHC-P: Human testis tissue. ICC/IF: HaCaT cells. Flow Cyt: HeLa cells.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.2
Concentration information loading...
PurityProtein G purified
Light chain typekappa
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.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration.|
|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.|
|ELISA||Use at an assay dependent concentration.|
|WB||1/2000. Predicted molecular weight: 85 kDa.|
|IP||Use at an assay dependent concentration.|
FunctionMolecular chaperone. Has ATPase activity.
Sequence similaritiesBelongs to the heat shock protein 90 family.
Cellular localizationCytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
- EL52 antibody
- epididymis luminal secretory protein 52 antibody
- Heat shock 86 kDa antibody
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.
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.
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.
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.
This product has been referenced in:
- 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). Read more (PubMed: 31370342) »
- Watanabe M et al. CD30 Induces Heat Shock Protein 90 and Signal Integration in Classic Hodgkin Lymphoma Cells. Am J Pathol 187:163-175 (2017). Read more (PubMed: 27870927) »