Stuff I'm Finding Kind of Hard to Remember Flashcards
Subset of Lab IgG Uses
To determine band/protein identity, quanitity and size on a gel (Western Blot)
To determine how much of a protein is present in a mixture (ELISA)
To determine the identity of a pathogen (not very definitive though for negative results)–Agglutination
To determine the antigens/antibodies in a sample (ouchterlony)
To localize proteins in a cell (fluoresence)
How is polyacrylamide made?
take acrylamide monomer, and react with ammonium persulfate and TEMED to make bisacrylamide (N,N’-methylene-bis-acrylamide)
The length of the polyacrylamide chains is dependent upon the concentration of the gel, while pore size is dependent upon the degree of cross-linking and length of the chains
ye
Relative mobility (Rf) measures the distance travelled by a protein relative to something else moving very fast (dye)
ye
Native gel are non-denaturing and they take much longer to run, because there are no extreme charges; also the charge depends on the pH used
ye
What are ampholytes?
Molecules that can buffer at their pI
On a 2D gel, there may be some spots with smears or streaks and this means it’s the same protein, but with some modification (acetylation, phosphorylation)
ye
What are the common modifications of plasmid isolation?
Resuspend and lyse in one step (dirtier, faster); addition of lysozyme to lysis buffer for gram (+); remove contaminating proteins using phenol/chlorogform extraction (slower, cleaner); resuspension of plasmid in buffer containing RNAse A
PCR Applications: Disease testing
Primers can be designed to give different sizes for function/non-functional genes; Primers can be designed to amplify ONLY functional/non-functional genes; Primers can be designed to give a product that is cut/not cut to reflect the presence of a diseased gene; PCR can amplify diseased genes for study (sequencing)
