Product nameAnti-Hsp90 antibody [D7a]
See all Hsp90 primary antibodies
DescriptionMouse monoclonal [D7a] to Hsp90
Tested applicationsSuitable for: WB, IP, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Rabbit, Chicken, Cow, Human, Pig
Full length native protein (purified) corresponding to Chicken Hsp90 alpha aa 1-728. Full length protein HSP90 purified from chicken brain
Database link: P11501
- HeLa (heat shocked) cell lysate; human Hsp90 alpha protein.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.2
Preservative: 0.09% Sodium azide
Constituents: 49% PBS, 50% Glycerol
Concentration information loading...
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.
|WB||1/500. Predicted molecular weight: 95 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionMolecular 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 similaritiesBelongs to the heat shock protein 90 family.
DomainThe TPR repeat-binding motif mediates interaction with TPR repeat-containing proteins like the co-chaperone STUB1.
S-nitrosylated; negatively regulates the ATPase activity and the activation of eNOS by HSP90AA1.
Cellular localizationCytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
- 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
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.
ab59459 staining Hsp90 alpha in the THP-1 monocyte/macrophage cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA, permeabilized with Triton X-100 0.1% and blocked with 5% Serum for 60 minutes at 20°C. Samples were incubated with primary antibody (1/200 1% BSA/5% Goat Serum) for 1 hour at 20°C. An Alexa Fluor®647-conjugated Goat anti-mouse IgG polyclonal(1/500) was used as the secondary antibody.
Ab59459 staining Hsp90 in Human DU145 cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with formaldehyde; permeabilized with 0.2% Triton X-100 in PBS and blocked with 1% Donkey serum in 0.1% PBST for 1 hour at 22°C. Samples were incubated in primary antibody at 1/50 dilution for 3 hours at 22°C. An Alexa Fluor® 488 Donkey anti-Mouse was used as the secondary antibody at 1/200 dilution.
Anti-Hsp90 antibody [D7a] (ab59459) at 1/1000 dilution + Rat tissue lysates
Predicted band size: 95 kDa
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.
Overlay histogram showing HeLa cells stained with ab59459 (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 (ab59459, 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 antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This product has been referenced in:
- Wang Y et al. Heat-shock protein 90a is involved in maintaining the stability of VP16 and VP16-mediated transactivation of a genes from herpes simplex virus-1. Mol Med 24:65 (2018). Read more (PubMed: 30577726) »
- Perez-Branguli F et al. Reverse Signaling by Semaphorin-6A Regulates Cellular Aggregation and Neuronal Morphology. PLoS One 11:e0158686 (2016). Read more (PubMed: 27392094) »