Recombinant Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)
![Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)](https://www.abcam.com/ps/products/222/ab222396/Images/ab222396-321041-anti-hif-2-alpha-antibody-epr19656-immunofluorescence.jpg "Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19656] to HIF-2-alpha - BSA and Azide free
- Suitable for: IP, WB, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free
See all HIF-2-alpha primary antibodies -
Description
Rabbit monoclonal [EPR19656] to HIF-2-alpha - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IP, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Whole cell lysate of HeLa cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h ; Whole cell lysate of HeLa cells treated with 100µM CoCl2 for 24h. ICC/IF: HeLa cells treated with DFO mesylate salt (1mM, 24h). IP: HeLa cell lysate.
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General notes
ab222396 is the carrier-free version of ab207607.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19656 -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222396 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 |
|---|---|---|
| IP |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 96 kDa).
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| ICC/IF |
Use at an assay dependent concentration.
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| Notes |
|---|
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IP
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 96 kDa). |
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ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Transcription factor involved in the induction of oxygen regulated genes. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of lung. May also play a role in the formation of the endothelium that gives rise to the blood brain barrier. Potent activator of the Tie-2 tyrosine kinase expression. Activation seems to require recruitment of transcriptional coactivators such as CREBPB and probably EP300. Interaction with redox regulatory protein APEX seems to activate CTAD. -
Tissue specificity
Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells. -
Involvement in disease
Defects in EPAS1 are the cause of erythrocytosis familial type 4 (ECYT4) [MIM:611783]. ECYT4 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated hemoglobin concentration and hematocrit, and normal platelet and leukocyte counts. -
Sequence similarities
Contains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 PAC (PAS-associated C-terminal) domain.
Contains 2 PAS (PER-ARNT-SIM) domains. -
Post-translational
modificationsIn normoxia, is probably hydroxylated on Pro-405 and Pro-531 by EGLN1/PHD1, EGLN2/PHD2 and/or EGLN3/PHD3. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.
In normoxia, is hydroxylated on Asn-847 by HIF1AN thus probably abrogating interaction with CREBBP and EP300 and preventing transcriptional activation.
Phosphorylated on multiple sites in the CTAD.
The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 2034 Human
- Omim: 603349 Human
- SwissProt: Q99814 Human
- Unigene: 468410 Human
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Alternative names
- Basic helix loop helix PAS protein MOP2 antibody
- Basic-helix-loop-helix-PAS protein MOP2 antibody
- bHLHe73 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, untreated or treated with deferoxamine mesylate salt (1mM, 24h), labeling HIF-2-alpha with ab207607 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on HeLa cells treated with deferoxamine mesylate salt (1mM, 24h).
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207607).
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![Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)](https://www.abcam.com/ps/products/222/ab222396/Images/ab222396-2-anti-hif-2-alpha-antibody-epr19656-immunoprecipitation-hela1.jpg)
Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)HIF-2-alpha was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell treated with 1mM DFO for 24h, cell lysate 10 µg with ab207607 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab207607 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate (10 µg) treated with 1mM DFO for 24h
Lane 2: ab207607 IP in DFO treated HeLa cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207607 in HeLa cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 75 seconds
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207607).
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![Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)](https://www.abcam.com/ps/products/222/ab222396/Images/ab222396-3-benefits-of-recombinant-antibodies.png)
Anti-HIF-2-alpha antibody [EPR19656] - BSA and Azide free (ab222396)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab222396 has not yet been referenced specifically in any publications.
