Anti-p53 antibody

• WB

Lab

Western blot - Anti-p53 antibody (AB131442)

Lanes 1 - 6 : Merged signal (red and green). Green - ab131442 observed at 53 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab131442 was shown to react with p53 in wild-type HAP1 cells in Western blot with loss of signal observed in TP53 knockout sample. Wild-type HAP1 and TP53 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab131442 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-p53 antibody (ab131442) at 1/1000 dilution

Lane 1:

Saos-2 (p53 null) cell lysate at 20 µg

Lane 2:

HL-60 (p53 null) cell lysate at 20 µg

Lane 3:

MCF7 (wildtype p53) cell lysate at 20 µg

Lane 4:

A431 (mutant p53) cell lysate at 20 µg

Lane 5:

Wild-type HAP1 cell lysate at 20 µg

Lane 6:

TP53 knockout HAP1 cell lysate at 20 µg

Predicted band size: 43 kDa

Observed band size: 53 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach cancer tissue labeling p53 with ab131442 at 1/200 dilution (overnight at 4°C). Heat mediated antigen retrieval was performed using Sodium citrate buffer pH 6 (>98°C, 20 minutes). Secondary antibody was used at 1/200 dilution for 30 minutes at room temperature.

Negative control : Secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling p53 with ab131442 at 1/200 (4°C, overnight). Cy3 labeled secondary antibody was diluted at 1/300 and incubated for 50 minutes at room temperature (red). Nuclei stained with DAPI (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemical analysis of paraffin-embedded human lung tissue labeling p53 with ab131442 at 1/200 (4°C, overnight). Cy3 labeled secondary antibody was diluted at 1/300 and incubated for 50 minutes at room temperature (red). Nuclei stained with DAPI (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue labeling p53 with ab131442 at 1/200 dilution (overnight at 4°C). Heat mediated antigen retrieval was performed using Sodium citrate buffer pH 6 (>98°C, 20 minutes). Secondary antibody was used at 1/200 dilution for 30 minutes at room temperature.

Negative control : Secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling p53 with ab131442 at 1/200 (4°C, overnight). Cy3 labeled secondary antibody was diluted at 1/300 and incubated for 50 minutes at room temperature (red). Nuclei stained with DAPI (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat spleen tissue labeling p53 with ab131442 at 1/200 dilution (overnight at 4°C). Heat mediated antigen retrieval was performed using Sodium citrate buffer pH 6 (>98°C, 20 minutes). Secondary antibody was used at 1/200 dilution for 30 minutes at room temperature.

Negative control : Secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling p53with ab131442 at 1/200 (4°C, overnight). Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20 min). Secondary antibody was diluted at 1/200 (RT, 30min).
Negative control was used by secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling p53with ab131442 at 1/200 (4°C, overnight). Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20 min). Secondary antibody was diluted at 1/200 (RT, 30min).
Negative control was used by secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody (AB131442)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse testis tissue labeling p53 with ab131442 at 1/200 dilution (overnight at 4°C). Heat mediated antigen retrieval was performed using Sodium citrate buffer pH 6 (>98°C, 20 minutes). Secondary antibody was used at 1/200 dilution for 30 minutes at room temperature.

Negative control : Secondary antibody only.

• WB

Lab

Western blot - Anti-p53 antibody (AB131442)

Lanes 1 - 4 : Merged signal (red and green). Green - ab131442 observed at 53 kDa. Red - loading control, ab8245, observed at 37 kDa.

Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab131442 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Other recommended antibodies to this target are ab32389 and ab26.

All lanes:

Western blot - Anti-p53 antibody (ab131442) at 1/500 dilution

Lane 1:

Wild type HAP1 whole cell lysate (20 µg)

Lane 2:

p53 knockout HAP1 whole cell lysate (20 µg)

Lane 3:

Saos2 (20 µg)

Lane 4:

A431 (20 µg)

Predicted band size: 43 kDa,53 kDa

false

• WB

Supplier Data

Western blot - Anti-p53 antibody (AB131442)

Loading control : Beta actin monoclonal antibody (5B7) at 1/5000 dilution followed by Dylight 680 secondary antibody at 1/10000 dilution (red).

Primary antibodies incubated overnight at 4°C.

Secondary antibodies incubated for 1 hour at 37°C.

All lanes:

Western blot - Anti-p53 antibody (ab131442) at 1/1000 dilution

All lanes:

Lysate from p53 knockout A431 (human epidermoid carcinoma cell line) cells

Secondary

All lanes:

Dylight 800 secondary antibody (green) at 1/10000 dilution

Predicted band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-p53 antibody (AB131442)

Western blot analysis showing p53 expression in wild-type and p53 shRNA knockout in rat PC-12 cells. Beta-actin served as the loading control.

This analysis shows the antibodies specificity due to detection in the endogenous p53 protein in wild-type but not in shRNA knockdown cell lysate.

The lower band may represent the physiologically degraded p53 protein

All lanes:

Western blot - Anti-p53 antibody (ab131442)

Lane 1:

Wild-type PC-12 (Rat adrenal gland pheochromocytoma cell line) cells

Lane 2:

p53 shRNA knockout PC-12 (Rat adrenal gland pheochromocytoma cell line) cells

Predicted band size: 43 kDa

false