ChIP assays were performed using the U-2 OS (human osteosarcoma cell line), TP53 polyclonal antibody (Cat # PAB14128) and optimized PCR primer sets for PCR. Chromatin sheared from 1x106 cells and 2 ug of antibody against TP53, or beads only were used per ChIP experiment. Figure shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Upper : recovery by TP53 of the TP21 promoter, a known target for TP53 (1), or beads only. Recovery of the TP21 promoter by the TP53 antibody is evident based on fluorescent qPCR analysis of immunoprecipitated DNA. Right : recovery of the GAPDH promoter (used as a negative control) by TP53 or beads only.
ELISA was performed using a serial dilution of TP53 polyclonal antibody (Cat # PAB14128), crude serum and flow through in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1 : 4,000.
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
p53 antigen,p53 transformation suppressor,p53 tumor suppressor,phosphoprotein p53,transformation-related protein 53