Introduction to gel migration or electrophoretic mobility experiments
Gel migration or electrophoretic mobility experiments (EMSA) can: Gel migration or electrophoretic mobility experiment The EMSA-electrophoretic mobility shift assay is a technique for studying the interaction of DNA-binding proteins with their associated DNA-binding sequences for qualitative and quantitative analysis. This technique was originally used to study DNA binding proteins and has been used to study the interaction of RNA binding proteins with specific RNA sequences. The purified protein and the crude cell extract are typically incubated with a 32P isotopically labeled DNA or RNA probe, and the complex and unbound probe are separated on non-denaturing polypropylene gel electrophoresis. DNA-complexes or RNA-complexes move slower than unbound probes. Isotopically labeled probes may be double-stranded or single-stranded depending on the binding protein being studied. When detecting a DNA binding protein such as a transcriptional regulatory factor, a purified protein, a partially purified protein, or a nuclear cell extract can be used. In the detection of RNA-binding proteins, purified or partially purified proteins may be used depending on the position of the RNA-binding protein of interest, and nuclear or cytoplasmic cell extracts may also be used. In competition experiments, DNA or RNA fragments and oligonucleotide fragments (specific) containing protein-binding sequences, and other unrelated fragments (non-specific) are used to determine the specificity of DNA or RNA-binding proteins. Specific binding is determined in the presence of competitive specific and non-specific fragments based on the characteristics and strength of the complex. Power Mints,Pvz Power Mints,Power Mints Candy,Best Power Mints Soar Biotech Co.,Ltd , https://www.xylitgum.com
(1) studying the interaction of DNA binding proteins and their associated DNA binding sequences;
(2) can be used for qualitative and quantitative analysis of DNA;
(3) For studying the interaction of RNA binding proteins with specific RNA sequences.