p53 Antibody (PSH03-95)

Immunoprecipitation: p53 Antibody (PSH03-95) [NBP3-32697]

Immunoprecipitation: p53 Antibody (PSH03-95) [NBP3-32697] - p53 was immunoprecipitated in 0.2mg NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate with NBP3-32697 at 2 ug/25 ul agarose. Western blot was performed from the immunoprecipitate using NBP3-32697 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate (input)
Lane 2: NBP3-32697 IP in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate
Lane 3: Rabbit IgG instead of NBP3-32697 in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes

Western Blot: p53 Antibody (PSH03-95) [NBP3-32697]

Western Blot: p53 Antibody (PSH03-95) [NBP3-32697] - Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (NBP3-32697) at 1/1,000 dilution.

Lane 1: RAW264.7 cell lysate
Lane 2: Neuro-2a cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 0.5uM doxorubicincell for 24 hours cell lysate

Lysates/proteins at 20 ug/Lane.

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 3 minutes 20 seconds;

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 (NBP3-32697) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/ Immunofluorescence: p53 Antibody (PSH03-95) [NBP3-32697]

Immunocytochemistry/ Immunofluorescence: p53 Antibody (PSH03-95) [NBP3-32697] - Immunocytochemistry analysis of NIH/3T3 cells treated with 0.5μM doxorubicin for 20 hours labeling p53 with Rabbit anti-p53 antibody (NBP3-32697) at 1/200 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-p53 antibody (NBP3-32697) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.