Recombinant Anti-p53 antibody [SP161] (ab227655)

Formalin-fixed, paraffin-embedded human stomach adenocarcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and TP53 knockout HAP1 cells stained with ab227655 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab227655) (1x10⁶ in 100 µl at 0.008 µg/ml) for 30 min at 22°C.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150177) was used at 1/2000 for 30 min at 22°C.

Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line TP53 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

Flow Cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling p53 with purified ab227655 at 1/20 dilution (7.5 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling p53 with purified ab227655 at 1/20 (7.25µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human bladder transitional cell carcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human lung squamous cell carcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human ovarian adenocarcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human B-cell lymphoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.