Transcription Factor Activity Assay Kits

Transcription Factor Activity Assays

The ELISA Genie Transcription Factor Activity Assay Kits are a premium range of Enzyme Linked Immunosorbent Assay (ELISA) kits used for the detection of active transcription factors in eukaryotic nuclear or cell lysates.

ELISA Genie Transcription Factor Activity Assay Kits allow scientists to perform high throughput screening of hundreds of samples in a convenient, efficient, and sensitive manner. This accelerates the journey from research to publication or development of a new drug.

Key ELISA Genie Transcription Factor Activity Assays


Features

  • Easy-to-use - Quicker, less complicated, and easier to achieve satisfactory results than common methods of detection of transcription factor binding to DNA elements and motifs.

  • Fast procedure - Significant reduction in runtime in comparison to common methods of detection of transcription factor binding to DNA elements and motifs - an ELISA Genie Transcription Factor Activity Assay can be completed in less than one day.

  • Safer - Eliminates the need for harmful radioactive labeling.

  • Accurate and reliable - High sensitivity and signal-to-noise ratio.

Contents

  • Binding Buffer
  • Primary Antibody Diluent
  • Primary Antibody
  • Nuclear Wash Buffer
  • HRP-conjugated Secondary Antibody
  • Cytoplasmic Extraction Buffer
  • Nuclear Extraction Buffer
  • Nuclear Lysate Positive Control
  • Wild-Type Consensus dsDNA Oligonucleotide
  • Mutant Consensus dsDNA Oligonucleotide
  • Ready-to-use Substrate
  • Wash Buffer
  • Stop Solution

Applications

  • Detection and qualitative analysis of activated transcription factors in nuclear and cell lysates.

  • Signal transduction pathway analysis.

  • Establishment of target candidates in drug development.

Principle

The ELISA Genie Transcription Factor Activity Assay (ELISA) Kit contains components necessary for detection of active transcription factors in eukaryotic nuclear or cell lysates. This particular immunoassay utilizes the qualitative technique of an indirect ELISA.

Streptavidin is bound to the immunoassay plate and specific biotinylated double stranded (dsDNA) oligonucleotides are then added to bind to the streptavidin via a high affinity biotin-streptavidin interaction.

After subsequent blocking of extraneous binding sites in each well, the sample containing the target of interest can be added. Primary antibody is added to bind activated transcription factors bound to the dsDNA oligonucleotide, which has been immobilized via the plate coated streptavidin.

A HRP-conjugated secondary antibody is added, which allows for specific binding to the Primary Antibody, and consequently colorimetric detection upon addition of the TMB (3, 3’, 5, 5’-Tetramethylbenzidine) substrate.

After addition of the substrate, a peroxidase catalysed reaction will produce a blue TMB Diimine product that is proportional to the target concentration in the sample. Colour development is quenched by addition of Stop Solution, or 2 N Sulfuric Acid, which turns the solution yellow.

The absorbance can then be read by a spectrophotometer at 450 nm and subsequently allowing for determination of the target concentration in the sample.

View a Transcription Factor Activity Assay Sample Protocol here.

Figure: Schematic representation of ELISA Genie Transcription Factor Activity Assay principle


Key ELISA Genie Transcription Factor Activity Assay Kits