Techniques in protein analysis Flashcards
1
Q
Involves the separation of soluble components in a solution
by specific differences in physical-chemical characteristics of
the different constituents.
A
Column chromatography
1
Q
Involves the separation of soluble components in a solution
by specific differences in physical-chemical characteristics of
the different constituents.
A
Column chromatography
2
Q
- Anionic detergent (-)
- Disrupt the structure of protein to be linear and binds most protein
- Sample run from negative charge to positive charge
A
SDS-PAGE
3
Q
Staining Method
A
Colorimetric
Fluorescence
4
Q
- Coomassie blue staining
- Zinc-reverse staining
- Silver staining
A
Colorimetric
5
Q
- CyeDyes
- Nile Red
- SyPro
- Epicocconone
A
Fluorescence
6
Q
- a laboratory method used to detect specific protein
molecules from among a mixture of proteins - The separated proteins are then transferred (blotted) to a
nitrocellulose or nylon sheet.
A
western blot
7
Q
- a laboratory method used to detect specific protein
molecules from among a mixture of proteins - The separated proteins are then transferred (blotted) to a
nitrocellulose or nylon sheet.
A
western blot
8
Q
Methods for Transferring Protein onto Membranes
A
- Diffusion
- Electroblotting
9
Q
COLORIMETRIC PROTEIN ASSAY TECHNIQUES
A
- Biuret Method
- Lowry Method
- Bicinchonic Acid Method
10
Q
- Enzyme-Linked ImmunoSorbent Assay
- common laboratory technique which is used to measure the
concentration of an analyte in a solution - uses the coupling of antigens and antibodies and
relies on the specificity and affinity of antibodies for antigens
A
ELISA
11
Q
- It uses a primary labeled antibody that react directly with the antigen
- It can be performed with the antigen that is directly immobilized on
assay plate - Not widely used bit common for immuno-histochemical staining of cells
and tissues
A
Direct ELISA
12
Q
- It utilizes a primary unlabeled antibody in conjunction with a labeled
secondary antibody - Secondary antibody has a specificity for primary antibody
- Antigen directly adsorbed onto solid phase is first incubated with patient
serum, and then with a labeled antibody specific for human
immunoglobulin
A
Indirect ELISA
13
Q
- Antigens like Tumor markers, hormones, serum proteins may be
determined - Antigens in the sample bind with the capture antibody and become
immobilized - The antibody of the enzyme conjugate bind with the immobilized antigen
to form a sandwich of Ab-Ag-Ab/ enzyme bound to microwell
A
Sandwich ELISA
14
Q
- used to determine the precise mass of peptides.
- The peptides are
immobilized in an organic matrix and then blasted with a laser, causing
them to be ejected in the form of an ionized gas. - The ionized peptides in
the gas are then accelerated in an electric field and separated
A
Matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF)
spectrometry
