Dieser p53 Antikörper ist konjugiert mit Alexa Fluor 647
Applikation
Intracellular Staining (ICS)
Marke
BD Phosflow™
Produktmerkmale
The p53 protein is critical to regulation of normal cell growth and proliferation and is a suppressor of tumor cell proliferation. Inactivation of p53 by a number of mechanisms, such as missense mutations or interaction with oncogenic viral or cellular proteins, can result in tumor progression. Mutations and/or allelic loss of the p53 gene are associated with a wide variety of human tumors. Known to have a role in transcriptional regulation, p53 suppresses various promoters containing TATA elements in an apparently sequence-independent fashion. p53 also binds to DNA in a sequence-specific manner via recognition of a 20-bp consensus-binding site. This interaction stimulates the expression of genes downstream of the p53 binding site. A number of genes that contain p53-binding sites have been identified, including MDM2, GADD45, and muscle creatine kinase. Post-translational acetylation of p53 enhances its DNA-binding activity. There are multiple factors that affect p53 acetylation, thereby modulating cellular proliferation and apoptosis. The L82-51 monoclonal antibody recognizes acetylated lysine 382 (acK382) in the C-terminal region of p53. Analysis of p53 (acK382) in lymphocytes. Human peripheral blood mononuclear cells (PBMC) were either treated with 0.4 μM Trichostatin A (Sigma, Cat. No. T8552) plus 0.4 μM Adriamycin (Doxorubicin hydrochloride, Sigma, Cat. No. D1515) for 24 hours (shaded histogram) or untreated (open histogram). The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-p53 (acK382). Lymphocytes were selected by scatter profile. Flow cytometry was performed on a BD FACSArray™ bioanalyzer system. The specificity of mAb L82-51 was confirmed by western blot analysis using unconjugated antibody on lysates from control (left blot) and Trichostatin A-plus-Adriamycin-treated (right blot) PBMC. p53 (acK382) is identified as a band of 53 kDa in the treated cells.
Optimal working dilution should be determined by the investigator.
Probenmenge
20 μL
Beschränkungen
Nur für Forschungszwecke einsetzbar
Buffer
Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.
Konservierungsmittel
Sodium azide
Vorsichtsmaßnahmen
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Lagerung
4 °C
Informationen zur Lagerung
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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