Electrophoresis Methods

Serum Protein Electrophoresis (SPE) (5)

1. Separate of proteins based on amount of negative charge

2. Stains: Coomassie blue, ponceau S, Silver stain

3. Cellulose acetate (8.6 pH)

4. Use densitometer to measure

5. Detect major differences in protein population

Immunofixation (IFE) (2)

1. Heavy and light chain composition of gamma proteins (Ig) first isolated via SPE via antibodies

2. Determine monoclonal gammopathy (M-Spike)

SDS-PAGE (5)

1. Separation of proteins based on size

2. Coated with SDS

3. Detection via protein stains and western blot + immunodetection

4. Acrylamide gel

5. Used for reasearch

Agarose Gel Electrophoresis (4)

1. Separation of nucleic acids based on size

2. Agarose gel

3. Detection: DNA stain (Coomassie blue, Ethidium bromide), southern blot + hybridization

4. Used for research

Isoelectric Focusing + 2D SDS-PAGE (4)

1. Separation of proteins first based on PI, followed by size

2. pH gradient gel

   1. Low pH left high pH right

3. Research

Capillary Electrophoresis

1. FAST separation of analytes

2. Electro-osmotic flow (EOF)

3. Hgb separation

Draw an image of a mass spec result. Indicate TWO purposes (there are 4)

1. Drug testing

2. ID unknown analytes (Forensics)

3. ID analyte fist separated vid chromatography (LC-MS and GC-MS)

4. Microbiology