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Fusion protein corresponding to Human HIF-1-alpha aa 432-528.
Database link: Q16665
Under normoxic conditions HIF-1 alpha has a short half-life. It is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% and upon stabilization under hypoxic conditions HIF-1 translocates to the nucleus. Therefore we recommend western blots using nuclear extracts and running Hypoxia treated samples as positive control (ab180880). Hypoxia can be induced with treatment using certain agents e.g. CoCl2 or DFO, etc. so proper sample preparation is critical.
Our Abpromise guarantee covers the use of ab463 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/100 - 1/500.|
|WB||1/500 - 1/1000.|
|Gel supershift assays||Use at an assay dependent concentration.|
|IHC-P||1/100 - 1/300. Antigen retrieval is not essential but may optimise staining.
Please check attached publications for more details.
|ELISA||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.|
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung tissue sections labeling HIF-1-alpha with ab463.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Dog breast cancer tissue sections labeling HIF-1-alpha with ab463 at 1/500. Tissue was fixed with formaldehyde and blocked with a peroxidase block for 5 minutes at 25°C; antigen retrieval was by heat mediation with EDTA buffer. Samples were incubated with primary antibody (1/500 in 1% BSA + PBS) for 30 minutes at 25°C. A biotin-conjugated Mouse anti-mouse IgG polyclonal was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling HIF-1-alpha with ab463.
This image is courtesy of an anonymous Abreview
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