The primary antibodies were: rabbit anti-human HIF-1α antibody (#3434T, 1:1,000), PTEN antibody (#9559T, 1:1,000), rabbit anti-human phospho-AKT antibody (#9271T, 1:1,000), and rabbit anti-human β-actin antibody (#4970T, 1:1,000) (all from Cell Signaling Technology). Horseradish peroxidase-conjugated (HRP) anti-rabbit antibodies (1:5,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA) were used as the secondary antibodies. The AKT inhibitor wortmannin was purchased from Sigma-Aldrich. The concentration used was 50 µM and the cells were treated for 12 h before further experiments. Cell lysates in 1X SDS loading buffer (60 mM Tris-HCl, pH 6.8; 2% SDS; 20% glycerol; 0.25% bromophenol blue; and 1.25% 2-mercaptoethanol) were incubated at 100°C for 10 min to facilitate sample loading for conventional western blot analysis. The relative protein levels were quantified using densitometry with a Gel-Pro Analyzer (Media Cybernetics, Rockville, MD, USA).
Rabbit anti human hif 1α antibody
Manufactured by Cell Signaling Technology
Sourced in United Kingdom
The Rabbit anti-human HIF-1α antibody is a primary antibody that specifically recognizes the human Hypoxia-Inducible Factor-1 alpha (HIF-1α) protein. HIF-1α is a transcription factor that plays a central role in the cellular response to hypoxia.
Automatically generated - may contain errors
Lab products found in correlation
Wortmannin, by Merck Group (1 mentions)
Pten antibody, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated hrp anti rabbit antibodies, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti human β actin antibody, by Cell Signaling Technology (1 mentions)
Gel pro analyzer, by Media Cybernetics (1 mentions)
Can get signal immunostain solution a, by Toyobo (1 mentions)
Bz 8000 confocal microscope, by Keyence (1 mentions)
2 protocols using rabbit anti human hif 1α antibody
1
Western Blot Analysis of HIF-1α, PTEN, and AKT
2
Immunohistochemical Analysis of Tumor Markers
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Formalin-fixed and paraffin-embedded tumor sections were pretreated with citrate buffer for 40 min at 95°C, quenched with 0.05% H2O2, and incubated overnight at 4°C with the following primary antibodies in Can Get Signal Immunostain Solution A (Toyobo): rabbit anti-human VEGF polyclonal antibody (1∶50) (Abcam, Cambridge, UK), rabbit anti-human HIF-1α antibody (1∶50) (Cell Signaling Technology), mouse anti-human MMP-2 antibody (1∶100) (Novocastra Laboratories, New-castle, UK) and mouse anti-human MMP-9 antibody (1∶100) (Novocastra Laboratories). Following this treatment, sections were incubated with horseradish peroxidase-conjugated goat anti-mouse IgG polyclonal antibody (Nichirei Bioscience, Tokyo, Japan) for 30 min at room temperature. Signals were developed as a brown reaction product using peroxidase substrate 3′,3′-diaminobenzidine (Nichirei Bioscience). The sections were counterstained with hematoxylin and examined with a BZ-8000 confocal microscope (Keyence, Osaka, Japan) [5] (link).
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