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LSBio DNA-Binding ELISA Kits
DNA-Binding ELISA Kits enable researchers to quickly and easily detect active transcription factors in eukaryotic nuclear 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.
Features:
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Kit Components:
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Features:
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Kit Components:
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DNA-Binding ELISA 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
| 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.
