CoraLite®594-conjugated P53 Monoclonal antibody

P53 Monoclonal Antibody for IF/ICC, FC (Intra)

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat

Applications

IF/ICC, FC (Intra)

Conjugate

CoraLite®594 Fluorescent Dye

CloneNo.

6C4B6

Cat no : CL594-60283

Synonyms

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

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Validation Data Gallery

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  IF Staining of A431 using CL594-60283

  Immunofluorescent analysis of (4% PFA) fixed A431 cells using CoraLite®594 P53 antibody (CL594-60283, Clone: 6C4B6 ) at dilution of 1:200.

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  FC experiment of HeLa using CL594-60283

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug CoraLite®594 Anti-Human P53 (CL594-60283, Clone:6C4B6) (red), or 0.4 ug Mouse IgG2b Isotype Control (CL594-66360-3, Clone: K11B8C4B5) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive IF/ICC detected in: A431 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• 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.

Product Information

CL594-60283 targets P53 in IF/ICC, FC (Intra) applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: P53 fusion protein Ag0698
• 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
• Conjugate: CoraLite®594 Fluorescent Dye
• Excitation/Emission Maxima Wavelengths: 588 nm / 604 nm
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

1.       What is p53?

P53 is a tumor suppressor gene that plays a role in maintaining genomic stability and controlling apoptosis. During the cell cycle, it can arrest cells at the G1/S checkpoint and activate DNA repair mechanisms. It is the most mutated gene in cancer. In unstressed cells, p53 usually exists at low levels in an inactive form, being bound to Mdm2.

2.       FAQs and p53

a.       I fail to detect p53 by western blotting

Basal levels of wild-type p53 in untreated cells can be low. Try to load more cell lysate and use a positive control - a lysate of cells treated with DNA-damaging agents should increase p53 levels.

b.      I fail to detect p53 in some cell lines by western blotting

Various p53 mutations are present in cancer cell types. If mutations cause truncations/deletions some monoclonal antibodies may no longer recognize mutated p53. You have more chances of detecting various p53 mutants with our polyclonal antibody.

c.       I can detect more than one band ~50 kDa size / different cell lines give bands at slightly different size

p53 is a subject of post-translational modifications (http://p53.free.fr/p53_info/p53_modifications.html) and more than one isoform may be expressed (http://p53.free.fr/p53_info/p53_isoforms.html). Also, it is possible that your cell line of interest expresses one allele with mutated p53 with altered molecular weight.