Nrf2 Recombinant Rabbit Monoclonal Antibody [PSH11-20]

☑ Cell treatment (CT)

Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA723302) at 1/2,000 dilution.

Lane 1: MEF cell lysate
Lane 2: MEF treated with 10μM MG-132 for 8 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 68 kDa
Observed band size: 100 kDa

Exposure time: 14 seconds; ECL: K1801;

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 (HA723302) at 1/2,000 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.

☑ Cell treatment (CT)

Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA723302) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 2μM MG-132 for 18 hours cell lysate
Lane 3: C6 cell lysate
Lane 4: C6 treated with 25μM MG-132 for 4 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 68 kDa
Observed band size: 100 kDa

Exposure time: 2 minutes; ECL: K1801;

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 (HA723302) at 1/2,000 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.

☑ Cell treatment (CT)

Immunocytochemistry analysis of MEF cells untreated / treated with 2μM MG-132 for 18 hours labeling Nrf2 with Rabbit anti-Nrf2 antibody (HA723302) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 (HA723302) 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.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

☑ Cell treatment (CT)

Immunocytochemistry analysis of HCT 116 cells untreated / treated with 25μM MG-132 for 4 hours labeling Nrf2 with Rabbit anti-Nrf2 antibody (HA723302) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 (HA723302) 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.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

☑ Cell treatment (CT)

Chromatin immunoprecipitations were performed with cross-linked chromatin from MEF cells treated with 10μM MG-132 for 8 hours with Nrf2 (HA723302) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.