p53 Transcription Factor Activity Assay Kit

• Cat.No.: Kit-2107
• Product Overview: The p53 Transcription Factor Activity Assay Kit is a non-radioactive assay for the quantitative measurement of specific transcription factor DNA binding activity of active p53. A biotin-labeled p53 response element DNA binds specifically to active p53 in nuclear extract and forms a complex. The amount of complex is detected by the p53 specific monoclonal antibody using BioVision"s High-Sensitivity Streptavidin Plate. A HRP-conjugated secondary antibody provides a sensitive colorimetric readout at 450 nm using TMB as substrate.

Specification

• Size: 100 assays
• Applications: Quantitative measurement of active p53 in cells or tissues for biomedical research and drug development
• Storage: Store the kit components, protected from light, according to specified storage condition. Briefly centrifuge small vials prior to opening. Read the entire protocol before performing the assay.Streptavidin Plate: Bring to room temperature before use. Reseal unused wells in the package and store at -20°C.Binding Buffer: Bring to room temperature before use. Prepare fresh Binding Buffer supplemented with 1 mM DTT by adding 2 µl of 1M DTT to 998 µl Binding Buffer. Prepare enough reagent per well (150 µl per each assay well). Use within 1 hour.Wash Buffer (10X): Bring to room temperature before use. Dilute with deionized water to prepare 1X working concentration. Prepare enough reagent per well (2 ml per each assay well).DTT, Capture Oligo, and Competitor Oligo: Thaw on ice before use.P53 Positive Control: Aliquot and store at -80°C. Thaw on ice before use. Avoid repeated freeze and thaw.Detection Ab: Dilute 100-fold with 1X Wash Buffer to prepare 1X detection Ab solution (100 µl per each well). Use within 1 hour.HRP Conjugate: Dilute 100-fold with 1X Wash Buffer to prepare 1X HRP Conjugates solution (100 µl per each well). Use within 1 hour.TMB Substrate, Stop Solution: Bring to room temperature before use. Ready to use.