Antibody Type: Recombinant Rabbit Monoclonal

Application: WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human

IgG

8Z908

Human

1. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-Nrf2 (Ser40)) monoclonal Antibody, Unconjugated (TMAB-01463) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
2. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-phospho-Nrf2 (Ser40) antibody (TMAB-01463)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-BF488
Dilution: 0.5 μg/Test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. 25 μg total protein per Lane of various lysates probed with phospho-Nrf2 (Ser40) monoclonal antibody, unconjugated (TMAB-01463) at 1:2000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.
4. Paraformaldehyde-fixed, paraffin embedded Human Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated (TMAB-01463) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated (TMAB-01463) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated (TMAB-01463) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Human Skin Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated (TMAB-01463) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated (TMAB-01463) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:50-200; IF: 1:100-500; FCM: 1μg/Test

Monoclonal

A synthesized peptide: human Nrf2 around the phosphorylation site of S40

Human

4780

Cardiac metabolism,Oxidative Stress,ChIP antibodies,Other factors,Mitochondrial transcription,Mitochondrial Biogenesis,Oxidative stress