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LSBio DNA-Binding ELISA Kits

DNA-Binding ELISA Kits
DNA-Binding ELISA Kits enable researchers to quickly and easily detect active transcription factors in eukaryotic nuclear extracts or cell lysates without the use of harmful radioactive reagents. Additionally our Phospho-DNA Binding kits can be used to study the effects of phosphorylation on transcription factor activation. Each kit is ready-to-use with all the necessary reagents and a clear, concise protocol that will step you through the process, from sample extraction to analysis of the results. DNA-Binding kits are available for both the detection of both phosphorylated and non-phosphorylated targets.


Alphabetical Listing


Features:

  • Ready-to-use kit includes all necessary reagents
  • Nuclear extraction protocol and reagents included
  • Phospho- and non-Phospho specific kits available
  • Detection of active transcription factors
  • Excellent sensitivity, specificity, and reproducibility
  • Built on standard 96-well microtiter plate format
  • 450 nm colorimetric detection

Kit Components:

  • 96-well microtiter plate
  • Anti-Phospho and Anti-Target Primary Antibodies
  • Nuclear Lysate Positive Control
  • Wild-Type Consensus dsDNA Oligonucleotide
  • Mutant Consensus dsDNA Oligonucleotide
  • Wash Buffers
  • Binding Buffer
  • Nuclear Wash Buffer
  • Nuclear and Cytoplasmic Extraction Buffers
  • HRP-Conjugated Secondary Antibody
  • TMB Substrate Solution
  • Stop Buffer
  • Plate Sealer Sheets

DNA-Binding ELISA Protocol

DNA-Binding ELISA Kits Protocol
1) The 96-well microtiter plate comes pre-bound with specific double-stranded (dsDNA) oligonucleotides.
2) Following a blocking step samples are added to each well and transcription factor binds to the oligonucleotides.
3) The wells are washed and primary antibody is added which binds activated transcription factor.
4) The wells are washed and a HRP-conjugated secondary antibody is added which binds to the primary antibody.
5) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow.


Phospho-Specific DNA-Binding ELISA Kit Protocol

Phospho-Specific DNA-Binding ELISA Kits Protocol
1) The 96-well microtiter plate comes pre-bound with specific double-stranded (dsDNA) oligonucleotides.
2) Following a blocking step samples are added to each well and phosphorylated transcription factor binds to the oligonucleotides.
3) The wells are washed and a phospho-specific primary antibody is added which binds activated transcription factor.
4) The wells are washed and a HRP-conjugated secondary antibody is added which binds to the primary antibody.
5) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow.


The plate can then be read in a spectrophotometer at a wavelength of 450 nm.
Nuclear Extract - DNA-Binding ELISA Kits TMB Substrate - DNA-Binding ELISA Kits