Recombinant Anti-Hsp90 antibody [EPR16621-67] (ab203126)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16621-67] to Hsp90
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Hsp90 antibody [EPR16621-67]
See all Hsp90 primary antibodies -
Description
Rabbit monoclonal [EPR16621-67] to Hsp90 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Hsp90 alpha recombinant protein fragment (GST-tag): aa533-732; Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724 ; Human fetal brain, fetal heart and fetal kidney lysates; Mouse and rat brain, heart and kidney lysates; HeLa, HEK-293, K562, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Human testis, Human lung cancer, mouse cerebral cortex and rat testis tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): Jurkat cells. IP: HeLa whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16621-67 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab203126 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 (Intra) |
1/350.
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ICC/IF |
1/200.
Methanol fixed cells. |
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IP |
1/100.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 85, 83 kDa).
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Notes |
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Flow Cyt (Intra)
1/350. |
ICC/IF
1/200. Methanol fixed cells. |
IP
1/100. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 85, 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 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: 3320 Human
- Entrez Gene: 3326 Human
- Entrez Gene: 15516 Mouse
- Entrez Gene: 15519 Mouse
- Entrez Gene: 299331 Rat
- Entrez Gene: 301252 Rat
- Omim: 140571 Human
- Omim: 140572 Human
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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All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : Rat kidney lysate
Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 85, 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunocytochemistry/Immunofluorescence analysis of 100% methanol-fixed NIH/3T3 (Mouse embryonic fibroblast) cells labeling Hsp90 alpha + beta with ab203126 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Control - PBS instead of primary antibody.
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Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on germ cells of Human testis is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of 100% methanol-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Hsp90 alpha + beta with ab203126 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Control - PBS instead of primary antibody.
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Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Hsp90 alpha + beta with ab203126 at 1/350 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
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Hsp90 alpha + beta was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab203126 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab203126 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab203126 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203126 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 3 seconds.
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Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on tumor cells of Human lung cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution
Lane 1 : Hsp90 alpha recombinant protein fragment (GST-tag): aa533-732
Lane 2 : Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 85, 83 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Human Hsp90 alpha and Hsp90 beta recombinant protein fragments were made in house.
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Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and nucleus staining on neuron of mouse cerebral cortex is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 85, 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on germ cells of rat testis is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Lanes 1 & 3-4 : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/100000 dilution
Lane 2 : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 85, 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (13)
ab203126 has been referenced in 13 publications.
- Rice MA et al. SU086, an inhibitor of HSP90, impairs glycolysis and represents a treatment strategy for advanced prostate cancer. Cell Rep Med 3:100502 (2022). PubMed: 35243415
- Zheng Z et al. Chronic Administration of COVID-19 Drugs Fluvoxamine and Lopinavir Shortens Action Potential Duration by Inhibiting the Human Ether-à-go-go-Related Gene and Cav1.2. Front Pharmacol 13:889713 (2022). PubMed: 35873575
- Huo J et al. Amphiphilic Aminated Derivatives of [60]Fullerene as Potent Inhibitors of Tumor Growth and Metastasis. Adv Sci (Weinh) 9:e2201541 (2022). PubMed: 36031401
- Chen YC et al. Induction of mitochondrial-dependent apoptosis by essential oil of Toona sinensis root through Akt, mTOR and NF-κB signalling pathways in human renal cell carcinoma cells. J Food Drug Anal 29:433-447 (2021). PubMed: 35696242
- Sun P et al. Hsp90 modulates human sperm capacitation via the Erk1/2 and p38 MAPK signaling pathways. Reprod Biol Endocrinol 19:39 (2021). PubMed: 33663544