Immunofluorescence analysis of p53 [pS15] was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ p53 [pS15] Recombinant Rabbit Monoclonal Antibody (700439) at 2 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear and cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 9-19 of P04637.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1 ul|
|Flow Cytometry (Flow)||0.5-5 µg/10^6 cells|
|Fluorescence Resonance Energy Transfer (FRET) (FRET)||Assay-dependent|
|Immunocytochemistry (ICC)||0.5-2 ug/ml|
|Immunofluorescence (IF)||0.5-2 ug/ml|
|Western Blot (WB)||0.1-0.5 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with chimpanzee, guinea pig, porcine, ovine, bovine, equine, feline, Rhesus monkey and Xenopus based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair, or apoptosis. p53 acetylation at Lys382 is required for recruitment of p300 to the p21 promoter, and is mediated by p300 and CBP acetyltransferases. Acetylation appears to play a positive role in the accumulation of p53 protein in stress responses, and inhibition of deacetylation stabilizes p53.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Automated quantitative analysis of tissue microarray of 443 patients with colorectal adenocarcinoma: low expression of Bcl-2 predicts poor survival.
700439 was used in immunohistochemistry - paraffin section to investigate Bcl2 expression in colorectal adenocarcinoma
|Nicholson AD,Guo X,Sullivan CA,Cha CH||Journal of the American College of Surgeons (219:977)||2014|
Profiling dose-dependent activation of p53-mediated signaling pathways by chemicals with distinct mechanisms of DNA damage.
700439 was used in western blot to develop high throughput methods to assess proof-of-concept in vitro-only risk assessments
|Clewell RA,Sun B,Adeleye Y,Carmichael P,Efremenko A,McMullen PD,Pendse S,Trask OJ,White A,Andersen ME||Toxicological sciences : an official journal of the Society of Toxicology (142:56)||2014|