Phospho-P53 (Ser46) Monoclonal antibody

Phospho-P53 (Ser46) Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

1D10A12

Cat no : 67900-1-Ig

Synonyms

TP53, P53, Tumor suppressor p53, tumor protein p53, Antigen NY-CO-13

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Validation Data Gallery

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  WB analysis using 67900-1-Ig

  Non-treated and Calyculin A treated HT-29 cells were subjected to SDS PAGE followed by western blot with 67900-1-Ig (Phospho-P53 (Ser46) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis using 67900-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 67900-1-Ig (Phospho-P53 (Ser46) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  IHC staining of human colon cancer using 67900-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 67900-1-Ig (Phospho-P53 (Ser46) antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HT-29 using 67900-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed etoposide treated HT-29 cells using Phospho-P53 (Ser46) antibody (67900-1-Ig, Clone: 1D10A12 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  FC experiment of HT-29 using 67900-1-Ig

  1X10^6 HT-29 cells were intracellularly stained with 0.4 ug Anti-Human Phospho-P53 (Ser46) (67900-1-Ig, Clone:1D10A12) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HT-29 cells, Calyculin A treated HT-29 cells, etoposide treated HT-29 cells, UV treated A431 cells, Calyculin A treated HEK-293 cells
• Positive IHC detected in: human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: etoposide treated HT-29 cells
• Positive FC (Intra) detected in: HT-29 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below

Product Information

67900-1-Ig targets Phospho-P53 (Ser46) in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: tumor protein p53
• Calculated Molecular Weight: 44 kDa
• Observed Molecular Weight: 53 kDa
• GenBank Accession Number: BC003596
• Gene Symbol: P53
• Gene ID (NCBI): 7157
• RRID: AB_2918656
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

P53 is a 53 kDa protein that is activated in response to alteration of normal cell homeostasis, including DNA damage, nutrient starvation, heat shock, virus infection, pH change, hypoxia, and oncogene activation. P53 maintains genetic stability by regulating different processes, such as cell-cycle arrest, DNA synthesis and repair, programmed cell death, and energy metabolism. In non-stressed conditions these proteins bind p53, ubiquitylate it and target it for degradation by the proteasome. In stressed conditions the function of the MdM2-MdM4 complex is blocked by phosphorylation, protein-binding events and/or enhanced degradation. (PMID: 19935675, PMID: 24379683)

Publications

Species	Application	Title
human	WB	J Cancer Res Clin Oncol; Urocortin-1 promotes colorectal cancer cell migration and proliferation and inhibits apoptosis via inhibition of the p53 signaling pathway; Authors - Xiaolan Guo

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Flow Cytometry Perm Buffer (10X)

PF00011-C