Lab skills 3 | Bright Minds Mentori

Immunoassay

Monoclonal Antibodies (MCA)

Stocks of antibodies with the same specificity to a particular antigen.

Immunoassay

Type 1 : Antigen on test well

A specific antigen on the test well is used to detect the presence of antibodies in a sample.

Sometimes a secondary MCA containing a chemical label is used to bind to primary MCA.

Type 2: MCA on Test well

A specific MCA on the test well is used to detect a particular antigen to detect the presence/concentration of antigen protein in the sample.

Stages of Immunoassay

It is important to wash away unbound molecules after adding MCA's OR antigens to avoid false positives by measuring unbound molecules.

Western Blotting

Identifies a protein from a complex mix using fluorescent labelled antibodies.

1. SDS Gel electrophoresis to separate proteins based on size.

2. The separated proteins are blotted onto a nylon membrane

3. Immunoassay: The protein is identified using specific antibodies that have reporter enzymes attached.

Microscopes

1. Bright Field

Low resolution allows the visualisation of

1. Whole organisms

2. Parts of organisms

3. Thin sections of dissected tissues

4. Individual cells

2. Fluorescent

Uses fluorescent labels to bind & visualise

specific molecules/structures within cells.

Higher resolution allows smaller structures

to be seen.