Friday, 30 September 2011

Promoter-Binding TF Profiling Assay I


To characterise transcription factors (TFs) that bind to a specific promoter or that regulate the expression of a specific gene via its upstream promoter, two common approaches may be applied.


  • Gel shift assay with DNA binding sites of TFs that are silico-identified within the promoter.
  • Removal or knockout the binding site(s) of a specific TF in order to measure whether the expression of a promoter-linked reporter is up or down regulated. Often, a series of reporter constructs with the promoter deletions or mutations need to be made as many binding sites of one or even a few TFs may be present within a promoter.
Signosis has developed a fast method to facilitate the characterisation of promoters through a revised TF activation array. This assay will help to test whether any of the selected 48 TFs bind to the promoter or not.

Benefits

The Signosis proprietary Promoter-Binding TF Profiling Array offers:
  • Multiplex Assay - A single assay permits the characterisation of the binding of 48 TFs to a specific promoter
  • Simple Procedures - Probe incubation, spin column separation, plate hybridisation, and HRP detection

Principle of the technology

In this assay, a PCR fragment containing the promoter of your interest is mixed with a set of 48 biotin-labelled oligos corresponding to 48 TFs along with an assayed sample. If unlabelled promoter DNA fragment contains a TF binding sequence, it will compete with the biotin-labelled oligo to bind to the TF in the sample, leading to a loss or reduction in biotin labelled TF/DNA complex formation. This will in turn result in either an absence or a significant reduction in signal. By comparing samples in the presence and absence of the competitor promoter DNA fragment, promoter-bound TFs can be identified.

For further information about this product, please follow this link.

1 comment:

  1. This is very good information.i think it's useful advice. really nice blog. keep it up!!!

    - multiplex assay

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