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Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA721432) at 1/1,000 dilution.
Lane 1: HeLa whole cell lysate
Lane 2: HeLa treated with 2μM MG-132 for 18 hours whole cell lysate
Lane 3: HCT 116 whole cell lysate
Lane 4: HCT 116 treated with 25μM MG-132 for 4 hours whole cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 110 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721432) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA721432) at 1/1,000 dilution.
Lane 1: NIH/3T3 cell lysate
Lane 2: NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 110 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721432) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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Immunocytochemistry analysis of HCT 116 cells labeling Nrf2 with Rabbit anti-Nrf2 antibody (HA721432) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nrf2 antibody (HA721432) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
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