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Human/Mouse/Rat Phospho-P53 (S15) and Total P53 ELISA

Human/Mouse/Rat Phospho-P53 (S15) and Total P53 ELISA

ArtNr	PEL-P53-S15-T-5
Hersteller	Raybiotech
Menge	5 Plate Kit
Quantity options	1 Plate Kit 2 Plate Kit 5 Plate Kit
Kategorie	Oncology ELISA ELISA & Immunoassays ELISA Kits Immunoassay By Organ Lung Cancer Bladder Cancer Colorectal Cancer Breast Cancer Testicular & Prostate Cancer
Typ	Elisa-Kit
Specific against	Human, Mouse, Rat
ECLASS 10.1	32160605
ECLASS 11.0	32160605
UNSPSC	41116126
Lieferbar
Specificity	The antibody pair provided in this kit recognizes human Phospho-P53 (pSer15) and total (pan) P53.
Species Detected	Human
Compatible Sample Types	Cell Lysates Tissue Lysates
Design Principle	Sandwich-based
Method of Detection	Colorimetric
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Gene Symbols	P53TP53
Kit Components	Pre- Coated 96-well Strip Microplate Wash Buffer Anti- Phospho Antibody Anti- Pan Antibody H R P- Conjugated Secondary Antibody Streptavidin- Conjugated H R P Assay Diluent T M B One- Step Substrate Stop Solution Lysis Buffer Positive Control Sample
Other Materials Required	Distilled or deionized water100 ml and 1 liter graduated cylinders Tubes to prepare sample dilutions Protease and Phosphatase inhibitors Precision pipettes to deliver 2 µl to 1 ml volumes Adjustable 1-25 ml pipettes for reagent preparation Benchtop rocker or shaker Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline	Prepare all reagents and samples as instructed in the manual. Add 100 µl of sample or positive control to each well. Incubate 2.5 h at R T or O/ N at 4 ° C. Add 100 µl of prepared primary antibody to each well. Incubate 1 h at R T. Add 100 µl of prepared 1 X H R P- Streptavidin to each well. Incubate 1 h at R T. Add 100 µl of T M B One- Step Substrate Reagent to each well. Incubate 30 min at R T. Add 50 µl of Stop Solution to each well. Read at 450 nm immediately.
Protein Name & Synonyms	Cellular tumor antigen p53 (Antigen NY-CO-13) (Phosphoprotein p53) (Tumor suppressor p53)
Pathway	Cell CycleDNA Damage
Introduction	RayBio® Phospho-P53 (S15) ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining phosphorylated P53 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human, mouse and rat phospho-P53. An anti-P53 (S15) antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated P53 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-P53 antibody is used to detect phosphorylated P53. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of P53 (S15). The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Positive Control	T47D cells were exposed to 50J/m2 of UV light followed by a 4 hours recovery period. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
UV irradiation of T47D Cell Line	T47D cells were untreated or exposed to 50J/m2 of UV light followed by a 4 hours recovery period before lysis. Cell lysates were analyzed using this phosphoELISA and Western Blot.

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