Recombinant Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [BL-95-1A2] to HIF-2-alpha
- Suitable for: ICC/IF, IP, ChIP-sequencing, ICC, IHC-P, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HIF-2-alpha antibody [BL-95-1A2]
See all HIF-2-alpha primary antibodies -
Description
Rabbit monoclonal [BL-95-1A2] to HIF-2-alpha -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, ChIP-sequencing, ICC, IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human HIF-2-alpha aa 400-450. The exact sequence is proprietary. NP_001421.2 and Gene ID 2034.
Database link: Q99814 -
General notes
This product is sold under License from Bethyl Laboratories, Inc.
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: 7.8
Preservative: 0.09% Sodium azide
Constituents: 98% Borate buffered saline, 0.1% BSA -
Concentration information loading...
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Purification notes
Recombinant antibody was purified from cell culture supernatant. -
Clonality
Monoclonal -
Clone number
BL-95-1A2 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab243861 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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ICC/IF |
1/100 - 1/500.
Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells. |
|
IP |
Use at an assay dependent concentration.
Use 5-20µl/mg lysate. |
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ChIP-sequencing |
Use a concentration of 10-40 - 30 µl/chromatin.
|
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ICC |
1/200 - 1/1000.
Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells. |
|
IHC-P |
1/200 - 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Antigen retrieval with citrate buffer pH 6.0 for 20 minutes using a pressure cooker and overnight incubations are recommended. |
|
WB |
1/1000.
|
Notes |
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ICC/IF
1/100 - 1/500. Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells. |
IP
Use at an assay dependent concentration. Use 5-20µl/mg lysate. |
ChIP-sequencing
Use a concentration of 10-40 - 30 µl/chromatin. |
ICC
1/200 - 1/1000. Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells. |
IHC-P
1/200 - 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. Antigen retrieval with citrate buffer pH 6.0 for 20 minutes using a pressure cooker and overnight incubations are recommended. |
WB
1/1000. |
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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Chromatin from subcutaneous human tumor 786-O cells was immunoprecipitated with anti-HIF-2-alpha antibody ab243861 and analyzed by
DNA sequencing. The figure illustrates the peak distribution of HIF-2-alpha binding within a 250 Kb region of chromosome 11 as detected
using anti-HIF-2-alpha antibody ab243861. ChIP-seq validation performed by Active Motif, Carlsbad, CA. -
Formaldehyde-fixed HepG2 (human liver hepatocellular carcinoma cell line) cells untreated (Left) and treated with CoCl2 (right), labeling HIF-2 alpha (Red) using ab243861 at 1/100 dilution. DyLight® 594-conjugated goat anti-rabbit IgG was used as the secondary antibody at 1/100 dilution. Phalloidin Alexa Fluor® 488conjugated (green) was used as the counterstain.
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Formalin-fixed, paraffin-embedded HepG2 (human liver hepatocellular carcinoma cell line) cells labeling HIF-2 alpha using ab243861 at 1/400 dilution in ICC/IF analysis. A HRP-conjugated goat-anti rabiit IgG was used as the secondary. DAB staining.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)Formalin-fixed, paraffin-embedded human renal cell carcinoma tissue stained for HIF-2 alpha using ab243861 at 1/400 dilution in immunohistochemical analysis. A HRP-conjugated goat anti-rabbit IgG was used as the secondary. DAB staining.
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HIF-2 alpha was immunoprecipitated from 1 mg HepG2 whole cell lysate with ab243861 at 20 µL per reaction. Western blot was performed on the immunoprecipitate using ab243861 at 1/1000 dilution.
Lane 1: ab243861 IP in HepG2 cell lysate treated with 200 µM CoCl2.
Lane 2: ab243861 IP in HepG2 whole cell lysate Mock treated.
Lane 3: Contol IgG in HepG2 cell lysate treated with 200 µM CoCl2.
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All lanes : Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861) at 1/1000 dilution
Lane 1 : HepG2 whole cell lysate treated with 200uM CoCl2
Lane 2 : HepG2 whole cell lysate mock treated
Lane 3 : HEK293T whole cell lysate treated with 200uM CoCl2
Lane 4 : HEK293T whole cell lysate mock treated
Lane 5 : 786-O whole cell lysate mock treated
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab243861 has been referenced in 5 publications.
- Lin JX et al. m6A methylation mediates LHPP acetylation as a tumour aerobic glycolysis suppressor to improve the prognosis of gastric cancer. Cell Death Dis 13:463 (2022). PubMed: 35568711
- Wang Z et al. Hypoxia-induced PLOD1 overexpression contributes to the malignant phenotype of glioblastoma via NF-?B signaling. Oncogene N/A:N/A (2021). PubMed: 33420370
- Yin R & Liu S SHARPIN regulates the development of clear cell renal cell carcinoma by promoting von Hippel-Lindau protein ubiquitination and degradation. Cancer Sci 112:4100-4111 (2021). PubMed: 34339558
- Liu X et al. HIF-1-regulated expression of calreticulin promotes breast tumorigenesis and progression through Wnt/β-catenin pathway activation. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 34706936
- Duan X & Qin G Notch inhibitor mitigates renal ischemia-reperfusion injury in diabetic rats. Mol Med Rep 21:583-588 (2020). PubMed: 31974606