Imprint® ChIP-Validated Monoclonal Anti-p53 (DO-1) antibody produced in mouse
SIGMA/SAB5100001 - clone DO-1, purified immunoglobulin
Synonym: Imprint® Monocolonal Anti-p53 antibody produced in mouse; Anti-LFS1; Anti-P53; Anti-TP53; Anti-TRP53; Anti-tumor protein p53
Product Type: Chemical
Immunoblotting Cell lysates of various cell lines were separated on SDS-PAGE and probed with 0.4 μg/mL Imprint® ChIP-Validated Monoclonal Anti-p53, Clone DO-1 (Cat. No. SAB5100001). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. A431 2. HEK-293T 3. RAJI
ChIP Validation ChIP Validation using Monoclonal Anti-p53 clone: DO-1 (Cat. No. SAB5100001). ChIP assays were performed using sonicated cross-linked chromatin from 1 × 107 MCF7 cells treated with 5-FU (Cat. No. F6627, 300 mM for 6 hours) according to Imprint® Ultra Chromatin Immunoprecipitation Kit (Cat. No. CHP2) using 2.3 μg Anti-p53 antibody/ChIP (Cat. No. SAB5100001) or 1 μg purified Mouse IgG (Cat. No. I5381). Successful enrichment of p53 associated DNA fragments was verified by qPCR analysis using primers for the promoters of the human p21 (CDKN1A/WAF1/CIP1) gene distal region (p21-D) and MDM2 gene. No enrichment was observed on an intergenic region on Chromosome 22 (Chr22) used as a negative control. The figure shows the p53 ChIP-qPCR enrichment on various specific targets plotted as a % of Input (100 % Input = total chromatin used for the ChIP reaction).
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | DO-1, monoclonal |
| conjugate | unconjugated |
| form | buffered aqueous solution |
| isotype | IgG2a |
| Quality Level | 200  |
| shipped in | dry ice |
| species reactivity | human |
| storage temp. | −20°C |
| technique(s) | ChIP: suitable |
| western blot: suitable | |
| UniProt accession no. | P04637  |
| Application: | Suitable for Chip and western blot. |
| Disclaimer: | This product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. |
| General description: | The optimal amount of antibody per ChIP assay should be empirically determined by the end-user depending upon the amount of chromatin used per reaction. We recommend testing 1-5 μg per ChIP assay. |
| General description: | The tumor suppressor protein p53 responds to diverse cellular stresses to regulate target genes that induce cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. Wildtype p53 protein is expressed at a low level in normal cells but often mutated and expressed at a high level in a variety of transformed cell lines, where it is believed to contribute to transformation and malignancy. p53 is a DNA-binding protein containing transcription activation, DNA-binding, and oligomerization domains. 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. |
| Immunogen: | recombinant human wild type p53 1-5. |
| Legal Information: | Imprint is a registered trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | View ChIP Validation Data for lot number: 032M4761 |
| Physical form: | Supplied as a solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide as a preservative. |
| Quality: | ChIP Validation |
| Storage and Stability: | For continuous use, store at 2-8°C for up to one month. For extended storage, freeze at -20°C in working aliquots. Repeated freezing and thawing or storage in “frost-free” freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |