Western blot, 0.1-0.5µg/ml Immunocytochemistry/Immunofluorescence, 5µg/ml ELISA, 0.1-0.5µg/ml
Flow Cytometry analysis of A431 cells using anti-Caspase 4 antibody. Overlay histogram showing A431 cells (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Caspase 4 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.
WB analysis of Caspase 4 using anti-Caspase 4 antibody.Lane 1:m
IF analysis of Caspase 4 using anti-Caspase 4 antibody. Caspase 4 was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-Caspase 4 Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Western blot analysis of Caspase 4 using anti-Caspase 4 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 50 ug of sample under reducing conditions. Lane 1: mouse liver tissue lysates, Lane 2: mouse testis tissue lysates, Lane 3: mouse thymus tissue lysates, Lane 4: mouse lung tissue lysates, Lane 5: mouse HEPA1-6 whole cell lysates, Lane 6: mouse NIH3T3 whole cell lysates, Lane 7: mouse SP20 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Caspase 4 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Caspase 4 at approximately 43KD. The expected band size for Caspase 4 is at 43KD.
Western blot analysis of Caspase 4 using anti-Caspase 4 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 50 ug of sample under reducing conditions. Lane 1: rat liver tissue lysates, Lane 2: rat testis tissue lysates, Lane 3: rat stomach tissue lysates, Lane 4: rat thymus tissue lysates, Lane 5: human COLO-320 whole cell lysates, Lane 6: human HepG2 whole cell lysates, Lane 7: human 22RV1 whole cell lysates, Lane 8: human PANC-1 whole cell lysates, Lane 9: human SGC-7901 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Caspase 4 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Caspase 4 at approximately 43KD. The expected band size for Caspase 4 is at 43KD.
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