Anti-p53 antibody [EPR20416-124]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-p53 antibody [EPR20416-124] (AB246223)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized , LLC1 (Mouse lung carcinoma) cell line labeling p53 with ab246223at 1/60 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabeled control (Cell without incubation with primary antibody and secondary antibody, Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/2000 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR20416-124] (AB246223)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LLC1 (mouse lung carcinoma) cells labeling p53 with ab246223 at 1/100 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077, green) at a 1/1000 dilution. Confocal image showing nuclear staining in LLC1 cells. An Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594, ab195889) was used as a counterstain at 1/200 dilution, shown in Red. The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

• IP

Unknown

Immunoprecipitation - Anti-p53 antibody [EPR20416-124] (AB246223)

p53 was immunoprecipitated from 0.35 mg LLC1 (Mouse lung carcinoma) whole cell lysate with ab246223 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab246223 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : LLC1 whole cell lysate (Input).
Lane 2 : ab246223 IP in LLC1 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246223 in LLC1 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-p53 antibody [EPR20416-124] (ab246223)

Predicted band size: 43 kDa

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• WB

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Western blot - Anti-p53 antibody [EPR20416-124] (AB246223)

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time :
Lane 1 : 26 seconds.
Lane 2 : 81 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 27702985).

All lanes:

Western blot - Anti-p53 antibody [EPR20416-124] (ab246223) at 1/1000 dilution

Lane 1:

LLC1 (mouse lung carcinoma) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-p53 antibody [EPR20416-124] (AB246223)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low or negative sample : normal brain (PMID : 32778161); normal liver (PMID : 10949934); NIH/3T3 (PMID : 10628835).

All lanes:

Western blot - Anti-p53 antibody [EPR20416-124] (ab246223) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse thymus tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 53 kDa,37 kDa

Observed band size: 53 kDa,37 kDa

false

Exposure time: 10s