Gel Shift (Gel) Methods, Techniques & Protocols

 

 

 

Definition: also called band shift assay, or electrophoretic mobility shift assay (EMSA), a method for detecting DNA-binding proteins.

General Methods & Techniques

Antigen-Antibody Specific Applications

DNA Binding Applications

RNA Binding Applications

Protein-DNA/Protein-RNA Interactions

Protein-DNA Complex

Virological Applications

Tumor, Disease & Diagnostic Applications

EMSA protocol - The EMSA, or Electrophoretic Mobility Shift Assay, is a method for studying DNA-protein interactions in vitro. DNA-protein complexes will migrate more slowly than unbound radioactive probe and are thus visualised by discrete bands of radioactivity near the top of an acrylamide gel image.

Band Shift Assay Protocol - The procedure described in this chapter for the determination of affinity constants and kinetic dissociation constants by band-shift assay refers to an ideal antibody fragment (e.g., a scFv or an Fab fragment) binding to a well-behaved protein antigen (pure, of well-defined oligomeric state, migrating as a single band in non-denaturing gel electrophoresis.

Use of P-FILM™ for Protein‑DNA Complex Identification - The gel shift, or electrophoretic mobility shift assay (EMSA) is used in the study of DNA – protein interactions. Traditionally, gel shift assays involve combining a protein (e.g., in vitro synthesized) or mixture of proteins (e.g., a nuclear extract) with a radioactively labeled DNA fragment containing a putative protein-binding site.

Gel Shift/Gel Super Shift Assays - The electrophoretic gel shift assay is used to detect sequence specific DNA-binding proteins present in nuclear extracts.