p53 Polyclonal Antibody

$Nuclear extracts of HeLa cells (40 \u03bcg) were analysed by Western blot using p53 Polyclonal Antibody (bs-53127R) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. Followed by secondary antibody incubation.$

Nuclear extracts of HeLa cells (40 \u03bcg) were analysed by Western blot using p53 Polyclonal Antibody (bs-53127R) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. Followed by secondary antibody incubation.

$ChIP assays were performed using human U2OS cells, treated with camptothecin, p53 Polyclonal Antibody (bs-53127R) and optimized PCR primer sets for QPCR. ChIP was performed on sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the p21 and GAS6 genes used as positive controls, and for GAPDH gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using human U2OS cells, treated with camptothecin, p53 Polyclonal Antibody (bs-53127R) and optimized PCR primer sets for QPCR. ChIP was performed on sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the p21 and GAS6 genes used as positive controls, and for GAPDH gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

$ChIP was performed on sheared chromatin from 4 million U2OS cells using 1 \u03bcg of the p53 Polyclonal Antibody (bs-53127R). The IP\u2019d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. The peak distribution along the X-chromosome and in 3 genomic regions of chromosome 6, 13 and 12, surrounding p21 (CDKN1A), GAS6 and MDM2, 3 known targets genes of p53.$

ChIP was performed on sheared chromatin from 4 million U2OS cells using 1 \u03bcg of the p53 Polyclonal Antibody (bs-53127R). The IP\u2019d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. The peak distribution along the X-chromosome and in 3 genomic regions of chromosome 6, 13 and 12, surrounding p21 (CDKN1A), GAS6 and MDM2, 3 known targets genes of p53.

To determine the titer of the antibody, an ELISA was performed using a serial dilution of p53 Polyclonal Antibody (bs-53127R), crude serum and flow through in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:308,000. Crude serum (blue), purified antibody (pink), flow through (yellow).

Applications

WB
ELISA
IF(ICC)
ChIP-seq

Reactivity

Human

Overview
Catalog #	bs-53127R
Product Name	p53 Polyclonal Antibody
Applications	WB, ELISA, IF(ICC), ChIP-seq
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against human p53 (tumor protein p53), using a KLH-conjugated synthetic peptide containing a sequence from the C-terminal part of the protein
Clonality	Polyclonal
Isotype	Not Determined
Concentration	2.5ug/ul
Purification	Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
Storage Buffer	PBS with 0.05% sodium azide.
Storage Condition	Store at -20°C for 12 months.
Target
Gene ID	7157
Swiss Prot	P04637
Synonyms	TP53, P53, TRP53, LSF1
Background	Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA-Mkln1. LincRNA-p21 participates in TP53-dependent transcriptional repression leading to apoptosis and seem to have to effect on cell-cycle regulation. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis. Regulates the circadian clock by repressing CLOCK-ARNTL/BMAL1-mediated transcriptional activation of PER2
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
IF(ICC)	1:50-200
ChIP-seq