Measuring Lymphocyte Transcription Factor Activity by ELISA
Adequate immune function requires the complex interaction between cells via a series of biochemical and molecular events, culminating in altered gene expression. Initiation of transcription by sequence‐specific DNA‐binding proteins known as transcription factors (TFs) is the principle point at which the expression of most genes is regulated. Thus, an understanding of nuclear events affected by environmental toxicants and their mechanisms of actions is critical to understanding toxic phenomena.
Colorimetric enzyme‐linked immunosorbent assay (ELISA)‐based procedures have been developed to detect specific transcription factor DNA‐binding activity in cell extracts. Up to 96 reactions can be performed in 3 to 4 hr. Extracts are added to the 96‐well plate precoated with a transcription factor DNA‐binding consensus sequence and detected with an antibody specific to the transcription factor of interest. In short, ELISA provides increased speed and throughput, and allows improved sensitivity and convenience over the traditional methods.
Keywords: Transcription Factor; DNA‐binding Activity; ELISA; Transcriptional Regulation
Basic Protocol 1: DNA‐Binding ELISA
Support Protocol 1: Coat Plates with DNA
Support Protocol 2: Prepare Nuclear Extracts
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