Anti-Hsp90 beta antibody [H90-10] (ab53497)
Key features and details
- Mouse monoclonal [H90-10] to Hsp90 beta
- Suitable for: Flow Cyt, IHC-P, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG2a
Overview
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Product name
Anti-Hsp90 beta antibody [H90-10]
See all Hsp90 beta primary antibodies -
Description
Mouse monoclonal [H90-10] to Hsp90 beta -
Host species
Mouse -
Specificity
Detects 90kDa. Detects HSP90 beta in all reactive species except in Chicken, where it detects both alpha and beta isoforms.
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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 beta.
Database link: P08238 -
Positive control
- WB: HEK-293T, Saos-2 and HL-60 cell lysates; Recombinant human Hsp90 beta protein. IHC-P: Human placenta, backskin and colon carcinoma tissue; mouse colon tissue. ICC/IF: HeLa 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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.2
Preservative: 0.09% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
H90-10 -
Isotype
IgG2a -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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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 ab53497 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 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
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IHC-P | (1) |
1/100.
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ICC/IF |
Use at an assay dependent concentration.
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WB | (2) |
1/2500. Predicted molecular weight: 83 kDa.
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Notes |
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Flow Cyt
Use 0.5µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
IHC-P
1/100. |
ICC/IF
Use at an assay dependent concentration. |
WB
1/2500. Predicted molecular weight: 83 kDa. |
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. -
Post-translational
modificationsUbiquitinated in the presence of STUB1-UBE2D1 complex (in vitro).
ISGylated.
S-nitrosylated; negatively regulates the ATPase activity. -
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: 3326 Human
- Entrez Gene: 15516 Mouse
- Omim: 140572 Human
- SwissProt: P08238 Human
- SwissProt: P11499 Mouse
- Unigene: 509736 Human
- Unigene: 2180 Mouse
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Alternative names
- 90 kda heat shock protein beta HSP90 beta antibody
- D6S182 antibody
- FLJ26984 antibody
see all
Images
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All lanes : Anti-Hsp90 beta antibody [H90-10] (ab53497) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HSP90AB1 knockout HEK-293T cell lysate
Lane 3 : Saos-2 cell lysate
Lane 4 : HL-60 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab53497 observed at 85 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab53497 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90AB1 knockout cell line ab266117 (HSP90AB1 knockout cell lysate ab257190). Wild-type and HSP90AB1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab53497 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Ab53497 staining Human normal placenta. Staining is localized to cytoplasmic compartment.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required. -
Immunohistochemistry analysis of formalin-fixed paraffin-embedded inflamed mouse colon using ab53497 at 1/10000. Primary antibody was incubated for 12 hours at 4°C. Secondary Antibody was a Biotin Goat Anti-Mouse at 1/2000 dilution incubated for 1 hour at room temperature. Counterstain was Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at room temperature. Magnification: 40x.
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ICC/IF image of ab53497 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53497, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Anti-Hsp90 beta antibody [H90-10] (ab53497) at 1 µg/ml +
Recombinant human Hsp90 beta protein (Active) (ab80033) at 0.1 µg
Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 83 kDa
Exposure time: 8 minutes -
Paraffin-embedded mouse backskin (epidermis) tissue fixed with Bouin's fixative, stained for Hsp90 beta using ab53497 at 1/100 dilution in immunohistochemical analysis. Primary antibody was incubated for 1 hour at room temperature. Secondary antibody was a FITC-conjugated goat anti-mouse (green) at 1/50 dilution ncubated for 1 hour at room temperature.
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Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for Hsp90 beta using ab53497 at 1/10000 in immunohistochemical analysis. Primary antibody was incubated for 12 hours at 4°C. Secondary antibody was an Alexa Fluor® 555 goat anti-mouse (red) at 1/5000 dilution incubated for 1 hour at room temperature.
40x magnification.
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ab53497 at 100,000 dilution staining Hsp90 in human colon cancer tissue section by immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). A antibody amplifier™ system was used for staining. A HRP-conjugated secondary antibody was used at 1/10 dilution
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ab53497 at 100,000 dilution staining Hsp90 beta in mouse colon tissue section by immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). A antibody amplifier™ system was used for staining. An Alexa Fluor® 568 conjugated secondary antibody was used at 1/10 dilution.
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All lanes : Anti-Hsp90 beta antibody [H90-10] (ab53497)
Lane 1 : Hsp90 beta protein
Lane 2 : Hsp90 alpha protein
Lysates/proteins at 2 µg per lane.
Predicted band size: 83 kDa -
Overlay histogram showing HeLa cells stained with ab53497 (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 (ab53497, 0.5µg/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 IgG2a [ICIGG2A] (ab91361, 1µg/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 (21)
ab53497 has been referenced in 21 publications.
- Khaledian B et al. Inhibition of heat shock protein 90 destabilizes receptor tyrosine kinase ROR1 in lung adenocarcinoma. Cancer Sci 112:1225-1234 (2021). PubMed: 33370472
- Liu Y et al. Proteomic Analysis Revealed the Characteristics of Key Proteins Involved in the Regulation of Inflammatory Response, Leukocyte Transendothelial Migration, Phagocytosis, and Immune Process during Early Lung Blast Injury. Oxid Med Cell Longev 2021:8899274 (2021). PubMed: 34007409
- Mottola G et al. Compromised thermal tolerance of cardiovascular capacity in upstream migrating Arctic char and brown trout-are hot summers threatening migrating salmonids? Conserv Physiol 8:coaa101 (2020). PubMed: 34868596
- Kovanicová Z et al. Cold Exposure Distinctively Modulates Parathyroid and Thyroid Hormones in Cold-Acclimatized and Non-Acclimatized Humans. Endocrinology 161:N/A (2020). PubMed: 32242612
- 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