|Validated Species:||Human, Mouse, Rat|
Data & Images
|Product Name||Anti-HIF-2-Alpha Antibody|
|Description||Rabbit IgG polyclonal antibody for Endothelial PAS domain-containing protein 1(EPAS1) detection. Tested with WB in Human;Mouse;Rat.|
|Cite This Product||Anti-HIF-2-Alpha Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1129-2)|
|Replacement Item||This antibody may replace the following items: sc-18473|sc-66847|sc-12690|sc-26160|sc-26159|sc-12688 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||A synthetic peptide corresponding to a sequence in the middle region of human HIF-2-alpha(282-301aa, HALDSENMTKSHQNLCTKGQ), identical to the related mouse and rat sequence.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Endothelial PAS domain-containing protein 1(EPAS-1)|
|Molecular Weight||96718 MW|
|Protein 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 (By similarity). .|
|Sequence Similarities||Contains 1 bHLH (basic helix-loop-helix) domain.|
|Subcellular Localization||Nucleus . Nucleus speckle . Colocalizes with HIF3A in the nucleus and speckles. .|
|Alternative Names||Endothelial PAS domain-containing protein 1;EPAS-1;Hypoxia-inducible factor 2-alpha;HIF-2-alpha;HIF2-alpha;Epas1;Hif2a;|
|Research Areas|||cardiovascular|hypoxia|hif| epigenetics and nuclear signaling|transcription|domain families|hlh / leucine zipper|hlh|cardiovascular/immune| cancer|invasion/microenvironment|chip'ing antibodies|cancer metabolism|response to hypoxia| metabolism|pathways and processes|metabolic signaling pathways|nucleotide metabolism|molecular processes|mitochondrial transcription|metabolism processes|types of disease||
Background for Endothelial PAS domain-containing protein 1(EPAS-1)
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-HIF-2-Alpha Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat|
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-HIF-2-Alpha Antibody Images
Click the images to enlarge.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat thymus tissue lysates,
Lane 2: rat testis tissue lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- HIF2A antigen affinity purified polyclonal antibody (Catalog # PA1129-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for HIF2A at approximately 110-120KD. The expected band size for HIF2A is at 96KD.
- Review by Christina Klinger
"We got bands as expected..."
--Christina Klinger, DEPT. OF TRANSLATIONAL AND REGENERATIVE MEDICINE, UNIVERSITY OF ARIZONA, MANAGER OF LABORATORY OPERATIONS
Applications Western Blot Sample Mouse lung lysates Detection SuperSignal West Femto Maximum Sensitivity Substrate
"We sought to see differences in expression of EPAS1 in protein from mouse lung samples. We selected this product because we have used other antibodies from Boster in the past with success. We got bands as expected, but they were not indicative of good conjugation as they were very light. We believe this is because we did not use the recommended Enhanced Chemiluminscent kit with anti-rabbit IgG (EK1002), so we will repeat the experiment with this chemi kit before concluding whether the antibody worked or not. This antibody should work using the chemiluminescent kit recommended...Heed the recommendation of the website and use the chemi kit suggested for high quality visualization."
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,