Transcription Factor Activity Assay Kits

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Semi-quantitive ELISA to detect activation of Transcription Factors by DNA probe coated microplate

The RayBio® Transcription Factor-Enzyme Linked Immunosorbent Assays are non-radioactive assays to semi-quantitatively measure active transcription factors in nuclear extracts and cell lysates. They offer an easy, speedy, sensitive and high-throughput method to detect the activation of transcription factors.

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ELISA kit

How It Works

TF-ELISA - How it works

Double stranded oligonucleotides are coated in a 96 well plate. These oligonucleotides specifically capture the target transcription factor contained in whole cell lysate or nuclear extracts after a short incubation. Subsequently, the primary antibody recognizes the target transcription factor-DNA complex in each well, and a HRP-conjugated secondary antibody is then used for detection. After washing away any unbound antibody, signal can be obtained easily through a colorimetric assay with a spectrophotometric plate reader at 450 nm. The specificity of the reaction between the target transcription factor and the DNA probe is additionally stringent because of the establishment of specific competitive DNA and non-specific competitive DNA probes in this reaction system.

Features

  • Specific transcription factor-DNA binding assay
  • Perfect alternative to EMSA
  • Easy to perform in an ELISA format
  • Non-radioactive assay
  • High throughput (96 well plate format)
  • Assay can be completed within 5 hours

Contents of Kit

  • 96-well Strip Microplate pre-coated with DNA probes
  • DNA Binding Buffer
  • Positive Control Sample
  • Specific and nonspecific Competitor DNA probe
  • Primary Antibody
  • Wash Buffer
  • Assay Reagent
  • DTT
  • HRP-conjugated Secondary Antibody
  • TMB One-Step Substrate Reagent
  • Stop Solution

Representative Data

Sandwich ELISA Standard Curve

Research Applications

  • Semi-quantitative detection of transcription factor-DNA complexes in nuclear extracts
  • Investigation of signal transduction pathways
  • Candidates scanning in drug development

Signaling Pathways