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Recombinant fragment corresponding to Human HIF-1 alpha (C terminal).
Database link: Q16665
HIF-1 alpha can be a difficult target to work with so we have compiled a summary of all the important information you need to know including useful tips. This can be found in the protocols tab or alternatively click here to download it.
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.
This product was changed from ascites to supernatant. Lot no’s high than GR206357-17 are from Tissue Culture Supernatant
Our Abpromise guarantee covers the use of ab113642 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||1/50 - 1/100.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||1/500 - 1/2000. Predicted molecular weight: 93 kDa.|
|IHC-P||1/200 - 1/1000.|
|ICC/IF||1/200 - 1/1000.|
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HIF-1 alpha (green) using ab113642 at 1/200 dilution. Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments labeled with Alexa Fluor® 555 phalloidin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver cancer tissue sections labeling HIF-1 alpha with ab113642 at a 1/200 dilution, followed by DAB staining.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung cancer tissue sections labeling HIF-1 alpha with ab113642 at a 1/200 dilution, followed by DAB staining.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach cancer tissue sections labeling HIF-1 alpha using ab113642 at a 1/200 dilution, followed by DAB staining.
Immunohistochemistry (Formalin-PFA-fixed paraffin-embedded sections) analysis human brain tumor tissue sections labeling HIF-1 alpha with ab113642 at a 1/200 dilution, followed by DAB staining.
Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab113642 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab113642, 1/100 dilution) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) ab96879 at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] ab91353, 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.
ELISA using Anti-HIF-1-alpha [1A3] antibody (ab113642).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"