Study of Transcription Factor Interaction with Chromatin by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0011-WXH)

Name: Study of Transcription Factor Interaction with Chromatin by Fluorescence recovery after photobleaching (FRAP)
SKU: STEM-MT-0011-WXH
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Introduction

Transcription factors are proteins involved in the process of converting, or transcribing, DNA into RNA. Transcription factors include a wide number of proteins, excluding RNA polymerase, that initiate and regulate the transcription of genes.
A typical transcription factor binds to DNA at a certain target sequence. Once it's bound, the transcription factor makes it either harder or easier for RNA polymerase to bind to the promoter of the gene.

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Principle Applications Procedure Materials

Principle

Fluorescence recovery after photobleaching (FRAP) is a microscopy technique capable of quantifying the mobility of molecules within cells. By exploiting the phenomenon of photobleaching, fluorescent mole- cules within a region of interest can be selectively and irreversibly 'turned off'. It is capable of quantifying the two-dimensional lateral diffusion of a molecularly thin film containing fluorescently labeled probes, or to examine single cells.

Applications

• Characterization of the mobility of individual lipid molecules within a cell membrane.
• Analysis of molecule diffusion within the cell
• Study of protein interaction partners, organelle continuity and protein trafficking.

Procedure

1. An initial fluorescence of fluorescent molecules is measured in the region of interest (ROI).
2. The fluorescent molecules are rapidly photobleached by focusing the high-intensity laser beam onto the defined area.
3. The exchange of bleached molecules with unbleached molecules from the surrounding region is followed over time using a low-intensity laser.