Anti-p53 antibody [EPR20416-120] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR20416-120] - BSA and Azide free (AB252380)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LLC1 (mouse lung carcinoma cell line) cells labeling p53 with ab246550 at 1/100 dilution followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in LLC1 cells is observed.

The nuclear counter stain is DAPI (blue). Tubulin is detected with an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246550).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-p53 antibody [EPR20416-120] - BSA and Azide free (AB252380)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized LLC1 (mouse lung carcinoma cell line) cell line labeling p53 with ab246550 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246550).

• IP

Unknown

Immunoprecipitation - Anti-p53 antibody [EPR20416-120] - BSA and Azide free (AB252380)

p53 was immunoprecipitated from 0.35 mg of LLC1 (mouse lung carcinoma cell line) whole cell lysate with ab246550 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab246550 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : LLC1 whole cell lysate 10 μg (Input).

Lane 2 : ab246550 IP in LLC1 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246550 in LLC1 whole cell lysate.

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

Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246550).

All lanes:

Immunoprecipitation - Anti-p53 antibody [EPR20416-120] (<a href='/en-us/products/primary-antibodies/p53-antibody-epr20416-120-ab246550'>ab246550</a>)

Predicted band size: 43 kDa

Observed band size: 53 kDa

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• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-p53 antibody [EPR20416-120] - BSA and Azide free (AB252380)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 U2OS cells treated with Etoposide (5μM 18h) and 5 µg of ab246550 [EPR20416-120]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246550).

• WB

Lab

Western blot - Anti-p53 antibody [EPR20416-120] - BSA and Azide free (AB252380)

This data was developed using ab246550, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) 1 : 1,000,000 dilution

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

All lanes:

Western blot - Anti-p53 antibody [EPR20416-120] (<a href='/en-us/products/primary-antibodies/p53-antibody-epr20416-120-ab246550'>ab246550</a>) 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

Observed band size: 53 kDa

false

Exposure time: 180s