Western blot, 0.25-0.5µg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5µg/ml, Human, Mouse, Rat Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Human ELISA, 0.1-0.5µg/ml, -
Flow Cytometry analysis of A431 cells using anti-HIF-1 alpha/HIF1A antibody. Overlay histogram showing A431 cells (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HIF-1 alpha/HIF1A Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
IHC analysis of HIF-1 alpha/HIF1A using anti-HIF-1 alpha/HIF1A antibody. HIF-1 alpha/HIF1A was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-HIF-1 alpha/HIF1A Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of HIF-1 alpha/HIF1A using anti-HIF-1 alpha/HIF1A antibody. HIF-1 alpha/HIF1A was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-HIF-1 alpha/HIF1A Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of HIF-1 alpha/HIF1A using anti-HIF-1 alpha/HIF1A antibody. HIF-1 alpha/HIF1A was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-HIF-1 alpha/HIF1A Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of HIF-1 alpha/HIF1A using anti-HIF-1 alpha/HIF1A antibody. HIF-1 alpha/HIF1A was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-HIF-1 alpha/HIF1A Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Western blot analysis of HIF-1 alpha/HIF1A using anti-HIF-1 alpha/HIF1A antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human A431 whole cell lysates, Lane 2: human HEPG2 whole cell lysates, Lane 3: human U20S whole cell lysates, Lane 4: rat PC-12 whole cell lysates, Lane 5: mouse thymus tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HIF-1 alpha/HIF1A antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour a
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