Tools for Analyzing Gene Expression Part 2

Vocabulary flashcards covering key concepts in Tools for Analyzing Gene Expression Part 2.

ELISA (Enzyme-Linked Immunosorbent Assay)

A quantitative method for detecting proteins using antibodies.

Protocol steps of ELISA

1. Coat well with antibody specific to target protein. 2. Add sample which may contain protein. 3. Add enzyme-linked antibody (HRP). 4. Add substrate (TMB), causing color change if protein is present.

EMSA (Electrophoretic Mobility Shift Assay)

A method for detecting DNA/protein or RNA/protein binding, where protein-bound nucleic acids migrate slower in a gel.

Supershift Assay

A technique that adds an antibody to the protein causing an even greater shift, confirming the protein's identity.

Co-Immunoprecipitation (Co-IP)

A method using antibodies to pull a protein from a sample along with its interacting partners.

Limitations of Co-IP

Weak interactions may break; identifying partner proteins requires additional antibodies.

SDS-PAGE

A technique often used in the analysis following Co-Immunoprecipitation, followed by Western blotting.

Secondary structure prediction

Can be predicted based on amino acid properties.

Tertiary structure prediction

More complex and less accurate compared to secondary structure prediction.

X-ray Crystallography

A method involving purification of protein, forming crystals, exposing to X-rays, and analyzing diffraction patterns to determine structure.

Limitations of X-ray Crystallography

Difficult crystal formation, requires high purity, and membrane proteins may not retain native shape.