Polycolonal and Monocolonal Antibodies
Terms
undefined, object
copy deck
- What type of antibody uses antigens to inoculate ( introduce an oily mixture to stimulate the immune system) ?
- Polycolonal Antibodies, Monocolonal Antibodies
- What is a soluble fraction from clotted blood in polycolonal antibodies?
- Serum
- Cross-reactivity is most associated with what?
- polycolonals and similar antigenic sites
- Which antibodies are associated witht he spleen?
- Monocolonal antibodies
- When the spleen and unfused cancer cells die, what lives with monoclonal antibodies?
- hybridoma cells live
- What does monoclonal mean?
- Made from one cell, one antibody.
- What are the first four steps of Westerns-immunoblot analysis?
-
1. seperate proteins by SDS-
Polyacrylamide Gel Electrophoresis (SDS PAGE),
2. electrophoretically transfer proteins from gel to special membrane paper,
3. block non-specific binding sites on membrane with inert protein solution,
4. Incubate membrane with primary Ab to protein of interest - What are the second four steps of Westerns-immunoblot analysis?
-
5. Wash excessively.
6. Incubate with secondary antibody that recognizes the primary Ab.
7. Wash excessively.
8. Develop for presence of secondary Ab. - What happens in SDS Polyacrylamide Gel Electrophoresis?
- proteins are seperated by molecular weight.
- What does incubating with secondary antibody that recognizes the primary Ab do?
- enhances detection. Secondary Ab are radioactive or enzyme linked.
- What are three uses of Antibodies (Ab)?
- Western Immunoblot Analysis, Immunoprecipitation, Immunofluorescence
- What are the first four steps of immuno-precipitation(fining proteins)?
-
1. Make extract from cells labeled with radioactivity,
2. Add primary Ab and incubate
3. Add secondary Ab (which binds the primary Ab) that is convalently attached to small beads and incubate
4. Centrifuge to collect beads - What are the second three steps of immuno-precipitation(finding proteins)?
-
5. release Ab/antigen complex with heat treatment and seperate on SDS page,
6. expose gel to film to identify size of protein that interacts with antibody (and other proteins that may interact with original antigen)
7. can also use non-radioactive cells and probe for different proteins in the pellet using the western immunoblot procedure - What are the first three steps of Immuno-fluorescence?
-
1. Grow cells on coverslip,
2. Fix cells on coverslip,
3. incubate cells with primary Ab, then wash - What are the second three steps of immunofluorescene?
-
4. incubate cells with secondary Ab linked to fluorescent molecule, wash
5. view cells with microscope with a uv light source
6. determine subcellular localization