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Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Human HIF3 alpha.
(Peptide available as ab20077.)
Our Abpromise guarantee covers the use of ab10134 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500. Detects a band of approximately 60 kDa (predicted molecular weight: 72 kDa).Can be blocked with Human HIF3 alpha peptide (ab20077).|
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
|IP||Use at an assay dependent concentration.|
ICC/IF image of ab10134 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab10134, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% milk before being incubated with ab10134 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. The band detected by ab10134 migrates at approximately 60-kDa. Although the predicted molecular weight of this protein is 72-kDa, the 60-kDa band has been observed in the literature. The intensity of this band has been shown to increase in cells exposed to hypoxic conditions (PMID:16775626).
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