Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free
See all p53 primary antibodies
Description

Rabbit monoclonal [EP155Y] to p53 (phospho S392) - BSA and Azide free
Host species

Rabbit
Specificity

This antibody is specific for p53 phosphorylated on Serine 392. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
Tested applications

Suitable for: IHC-P, WB, Dot blot, IPmore details
Unsuitable for: ICC/IF
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
General notes
Ab239211 is the carrier-free version of ab33889. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab239211 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP155Y
Isotype

IgG
Research areas

Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (purified)

Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates, then the membrane was incubated with alkaline phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 44 kDa

This data was developed using ab33889, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

This data was developed using ab33889, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling p53 with Purified ab33889 at 1:250 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (purified)

Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates, then the membrane was incubated with alkaline phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 44 kDa

This data was developed using ab33889, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (purified)

Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates, then the membrane was incubated with alkaline phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 44 kDa
Observed band size: 53 kDa
why is the actual band size different from the predicted?

This data was developed using ab33889, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

This data was developed using ab33889, the same antibody clone in a different buffer formulation.
ab33889 (purified) at 1:20 dilution (0.6μg) immunoprecipitating p53 in 293T whole cell lysate.

Lane 1 (input): 293T (Human embryonic kidney epithelial cell) whole cell lysate, 10μg
Lane 2 (+): ab33889 & 293T whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab33889 in 293T whole cell lysate.

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/2000 dilution (purified)

Lane 1 : A431 whole cell lysate
Lane 2 : A431 treated with 1Î¼g/ml doxorubicin for 24 hours whole cell lysate
Lane 3 : A431 treated with 1Î¼g/ml doxorubicin for 24 hours whole cell lysate, the membrane was incubated with alkaline phosphatase.

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 44 kDa
Observed band size: 53 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

This data was developed using ab33889, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

This data was developed using ab33889, the same antibody clone in a different buffer formulation.
Unpurified ab33889, at a 1/100 dilution, staining p53 in paraffin embedded human prostate adenocarcinoma tissue by Immunohistochemistry.
Dot Blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

This data was developed using ab33889, the same antibody clone in a different buffer formulation.Dot blot analysis of p53 (pS392) peptide (Lane 1) and p53 non-phospho peptide (Lane 2) labelling p53 (pS392) with unpurified ab33889 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000. Blocking and dilution buffer: 5% NFDM/TBST. Exposure time: 3 minutes.
Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/1000 dilution (unpurified)

Lane 1 : HEK-293 whole cell lysate - untreated
Lane 2 : HEK-293 whole cell lysate - treated with Alkaline Phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

Predicted band size: 44 kDa
Observed band size: 53 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

This data was developed using ab33889, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

This data was developed using ab33889, the same antibody clone in a different buffer formulation.

Lanes 1 and 5: Extract of Hek293T incubated with etoposide (7.5 µg).

Lanes 2 and 6: Lambda phosphatase (400 times-diluted)-treated extract of Hek293T incubated with etoposide (7.5 µg).

Lanes 3 and 7: Lambda phosphatase (100 times-diluted)-treated extract of Hek293T incubated with etoposide (7.5 µg).

Lanes 4 and 8: Lambda phosphatase (25 times-diluted)-treated extract of Hek293T incubated with etoposide (7.5 µg).

SDS PAGE performed under reducing conditions (100mM DTT, sample heated at 50°C).

Primary:

Lanes 1-4: Anti p53 (phosphoS392) antibody (ab33889, unpurified) at 1/2000 dilution.

Lanes 5-8: Anti p53 antibody (ab1101) at 1/2500 dilution.

Secondary:

Lanes 1-4: HRP-conjugated goat anti-rabbit IgG (H&L) at 1/10000.

Lanes 5-8: HRP-conjugated goat anti-mouse IgG (H&L) at 1/10000.

Blocked in 5% milk in PBS for 3 hours at room temperature.

Incubated with the primary antibody in 5% BSA + 50mM Tris pH 7.5 + 0.05% Tween-20 overnight at 4°C.

Incubated with the secondary antibody in blocking buffer for 2 hours at room temperature.
Western blot - Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)

All lanes : Anti-p53 (phospho S392) antibody [EP155Y] (ab33889) at 1/500 dilution (unpurified)

Lane 1 : MCF7 cell lysate untreated
Lane 2 : MCF7 cell lysate treated with 5 ug/ml Actinomycin for 3hrs.
Lane 3 : MCF7 cell lysate treated with 5 ug/ml Actinomycin for 6hrs.
Lane 4 : MCF7 cell lysate treated with 5 ug/ml Actinomycin for 18hrs.

Predicted band size: 44 kDa
Observed band size: 53 kDa why is the actual band size different from the predicted?

This data was developed using ab33889, the same antibody clone in a different buffer formulation.
Anti-p53 (phospho S392) antibody [EP155Y] - BSA and Azide free (ab239211)