TP53 monoclonal antibody, clone PAb421
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Specification
Product Description
Rabbit recombinant monoclonal antibody raised against TP53.
Antibody Species
Rabbit
Immunogen
Originate antibody is raised against synthetic peptide corresponding to amino acids 371-380 of human TP53
Reactivity
Human
Specificity
This antibody reacts to the mammalian mutant and wild-type TP53 protein and reacts with an epitope between amino acid residues 370 and 378 (near the C-terminus of the protein), corresponding to the sequence "KKGQSTSRHK".
Form
Liquid
Purification
Protein A affinity purified
Isotype
Rabbit IgG, kappa
Recommend Usage
Flow Cytometry
Immunofluorescence
Western Blot
The optimal working dilution should be determined by the end user.Storage Buffer
In PBS (0.02% Proclin 300)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing. -
Applications
Western Blot (Cell lysate)
Western blot analysis of TP53 monoclonal antibody, clone PAb421 (Cat # RAB01077).
A431 cell nuclear (A) and cytoplasmic (B) extract (35 ug protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit antibody (Cat # RAB01077) at 0.1 ug/mL before detection by an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for TP53 is 43.7 kDa, though due to the high number of proline residues in this protein runs at a size of ~53kDa (c.f. Ziemer et al., PMID: 7107651). Cat # RAB01077 successfully detected both human nuclear and cytoplasmic TP53.Immunofluorescence
Immunofluorescence staining of fixed A431 with TP53 monoclonal antibody, clone PAb421 (Cat # RAB01077).
Immunofluorescence analysis of paraformaldehyde fixed A431 cells, permeabilized with 0.15% Triton and stained with the chimeric rabbit IgG antibody (Cat # RAB01077) at 10 ug/mL for 1h followed by Alexa Fluor® 488 secondary antibody (1 ug/mL), showing nuclear staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Cat # RAB01077, DAPI, merged channels and a negative control. The negative control was stained with unimmunized rabbit IgG followed by Alexa Fluor® 488 secondary antibody.Flow Cytometry
Flow-cytometry using TP53 monoclonal antibody, clone PAb421 (Cat # RAB01077).
Jurkat cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric antibody (Cat # Ab00142, blue line) at a concentration of 10 ug/mL for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody at 2 ug/mL and cells analyzed on a FACSCanto flow-cytometer. -
Gene Info — TP53
Entrez GeneID
7157Protein Accession#
P04637Gene Name
TP53
Gene Alias
FLJ92943, LFS1, TRP53, p53
Gene Description
tumor protein p53
Gene Ontology
HyperlinkGene Summary
This gene encodes tumor protein p53, which responds to diverse cellular stresses to regulate target genes that induce cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. p53 protein is expressed at low level in normal cells and at a high level in a variety of transformed cell lines, where it's believed to contribute to transformation and malignancy. p53 is a DNA-binding protein containing transcription activation, DNA-binding, and oligomerization domains. It is postulated to bind to a p53-binding site and activate expression of downstream genes that inhibit growth and/or invasion, and thus function as a tumor suppressor. Mutants of p53 that frequently occur in a number of different human cancers fail to bind the consensus DNA binding site, and hence cause the loss of tumor suppressor activity. Alterations of this gene occur not only as somatic mutations in human malignancies, but also as germline mutations in some cancer-prone families with Li-Fraumeni syndrome. Multiple p53 variants due to alternative promoters and multiple alternative splicing have been found. These variants encode distinct isoforms, which can regulate p53 transcriptional activity. [provided by RefSeq
Other Designations
p53 antigen|p53 transformation suppressor|p53 tumor suppressor|phosphoprotein p53|transformation-related protein 53
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