One million bone marrow-derived macrophages were dispensed into wells of a 48-well tissue culture plate and incubated with medium alone or with 5 x106 Histoplasma capsulatum yeast cells or with 300 mg/ml of cobalt chloride (CoCl2) for 24 h at 37°C in 5% CO2. Western blot analysis of HIF-2 alpha was performed by loading 150 ug whole cell lysate in reducing sample buffer onto a 4-20% gradient acrylamide gel. Proteins were transferred to nitrocellulose.HIF-2 alpha was detected at approximately 97 kDa using a HRP conjugated HIF-2 alpha Monoclonal Antibody (ep190b) (Product # MA5-16023) at a dilution of 1:500 overnight at 4°C. Chemiluminescent detection was performed.
|Tested species reactivity||Hamster, Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Human HIF-2 alpha amino acids 535-631.|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||1.0 µg|
|Immunohistochemistry (Paraffin) (IHC (P))||1:150-1:300|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA5-16023 detects HIF-2 alpha in WB application with human samples.
Suggested positive control: antigen standard for EPAS1 (transient overexpression lysate).
Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease.
HIF-2 alpha is predominantly expressed in highly vascularized tissues of adult humans and endothelial cells of the embryonic and adult mouse, whereas HIF-1 alpha functions primarily in extravascular tissues. HIF-2 alpha is also a potent activator of the tie-2 gene, which is known to be selectively expressed in endothelial cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.