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IMCH (Fach) / Techniques (Lektion)
- Grabar and Williams Method · combination of electrophoresis of proteins (antigens) and diffusion of antibodies · 1. Step: electrophoretic separation AG-mixture · 2. Step: AB-mixture in channel pipeted à diffusion à precipitate bows
- RID Radial Immunodiffusion Gel with Antibody AG is pipeted into holes --> diffusion --> ringdiameter is measuerd
- Laurell Immune-electrophoersis Rockets · electrophoresis of AG in gel with monospecific AB · Quantitative: peakhight à amount of AG · · (basic pH no electrophoresic movement of AB) · during electr. à more binding of antibodies and building of precipitate at equivalence point
- Crossed immune-elctrophoresis 2D (Clarke & Freeman) · 1st dimension: AGs are separated and deposit at isoelectric point à cutted off · 2nd dimension: electroph. In antibody-containing gel à Immunoprecipitation · each precipitate representing one antigen · position dependent on amount of protein and antibody in gel à quantification
- Tandem crossed immune-electrophoresis · Second purified protein AG àidentification and quantification of AG in 2nd dimension
- Fused Rocket · Fractions from chromatographie in zick zack holes of gel with AB · Interdiffusion · electrophoresis · one-dimensional quantitative immunoelectrophoresis
- Crossed electrophoresis with intermediate gel containing Antiserum 1. gel with unknown sample 2. intermediate gel containing specific antiserum 3. gel containing polyvalent antiserum
- Western Blotting 1. AG-mixture electrophorized in SDS-polyacrylamide gel 2. Vertically electric field transfer to Membrane 3. Bind AG of interest with enzyme-linked ABs
- Immunfixation: 1. Electrophorese 2. Fixation 3. AB inkubated à preciptation 4. Coloured
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