1: Techniques in Protein Analysis (FINALS) Flashcards
→ The most abundant macromolecules in biological system
→ polymers comprising amino acids that are linked by peptide bonds
→ are diverse in their chemical and physical properties
→ serve as antibodies, enzymes, messengers, structural components, transport, transport or storage molecule
Proteins
Proteins are polymers comprise of what?
Amino acids
Proteins are amino acids linked by what?
Peptide bonds
T or F
many diseases can be distinguished from others based on the type of proteins
T
These proteins can be detected in the blood of the patient
Biomarkers
A measurable substance in the blood whose presence is indicative of
diseases or environmental exposure
Biomarkers
When can proteins be further purified? (2 conditions needed)
- After cells are broken
- Cell extracts are released
The most common method for purifying proteins from other protein molecules with a given sample
Column Chromatography
Involves the separation of soluble components in a solution by specific differences in physical- chemical characteristics of the different constituents
Column Chromatography
T or F
Separation of soluble components in solution through column chromatography happens due to the similarities of the physical-chemical characteristics
F (DIFFERENCES of the physical-chemical characteristics)
Column Chromatography uses what equipment?
glass or plastic tube with resin
Familiarize the method of column chromatography
- Protein sample is applied to the top of the column
- Buffer solution is flowed continuously through the column
- Proteins in sample migrate through column
In the Column Chromatography method, proteins in sample migrate at different rate due to what factors? (clue: tatlo ‘to)
- Nature
- Physical properties
- Chemical properties
T or F
Proteins in the sample migrate through the column at different rate depending on the nature of the matrix and the physical and chemical properties of the protein.
T
What are the 3 types of Column Chromatography method?
- Gel/Gel Permeation/
Gel Filtration/Size Exclusion - Ion Exchange Chromatography
- Affinity Chromatography
3 Types of Column Chromatography:
In this type of column chromatography, the resin bead has many tiny pores like a whiffle ball
Gel/Gel Permeation/
Gel Filtration/Size Exclusion
3 Types of Column Chromatography:
In this type of column chromatography, it separates molecules based on difference in their size and shape.
Gel/Gel Permeation/
Gel Filtration/Size Exclusion
Gel/Gel Permeation/
Gel Filtration/Size Exclusion Type of Column Chromatography:
Which are more likely to go through the pore of the matrix, the SMALL or LARGE molecules?
Small molecules
Gel/Gel Permeation/
Gel Filtration/Size Exclusion Type of Column Chromatography:
T or F
Small molecules move through the column more quickly since they cannot fit into the beads. - They are excluded from entering the pores of the beads
F (LARGE MOLECULES move through the column more quickly since they cannot fit into the beads. - They are excluded from entering the pores of the beads)
T or F
In Gel/Gel Permeation/
Gel Filtration/Size Exclusion, the first size of molecules to be eluted/separated are the large molecules
T
3 Types of Column Chromatography:
In this type of column chromatography, it uses resin to separate proteins according to their surface charges
Ion Exchange Chromatography
T or F
The resin in Ion Exchange Chromatography contains only positively charged chemical groups
F (contains EITHER POSITIVE OR NEGATIVELY CHARGED CHEMICAL GROUPS)
3 Types of Column Chromatography: Ion Exchange Chromatography
The concept to remember in Ion Exchange Chromatography?
Opposite charges attract
3 Types of Column Chromatography: Ion Exchange Chromatography
Anion-exchange/Resins with positively charged groups resins attract what?
negatively charged solute