Anti-p53 antibody [SP161] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; and sodium azide (ab227655)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227655).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab227655).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

Intracellular Flow Cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling p53 with purified ab227655 at 1/20 dilution (7.5μg/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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227655).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab227655).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; and sodium azide (ab227655)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab227655).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; and sodium azide (ab227655)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; and sodium azide (ab227655)

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-p53 antibody [SP161] - BSA and Azide free (AB245740)

This data was developed using the same antibody clone in a different buffer formulation (ab227655).

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-Rabbit IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150081) 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 gave a positive signal in TP53 HAP1 knockout cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.