PROTEIN ISOLATION, PURIFICATION, AND QUANTIFICATION

Question 1

is a complex procedure that involves
extracting a specific protein or a group
of proteins from a mixture containing
various cellular components.

Answer 1

Protein Isolation

Question 2

PROTEIN SOLATION TECHNIQUES
1.
2.

Answer 2

Cell Lysis
Centrifugation

Question 3

process is designed to remove unwanted contaminants such as other proteins, nucleic acids, lipids, and small molecules, resulting in a highly pure protein sample.

Answer 3

Protein Purification

Question 4

3 techniques in protein purification

Answer 4

1. Chromatography
2. Electrophoresis
3. Filtration

Question 5

It involves passing the protein mixture
through a column containing a stationary
phase, which interacts with the proteins
based on their specific characteristics such
as size, charge, or affinity.

Answer 5

Chromatography

Question 6

It involves applying an electric field
to a gel matrix containing the sample
mixture. The charged molecules in
the sample move through the gel at
different rates based on their size and
charge, resulting in separation.

Answer 6

Electrophoresis

Question 7

techniques utilize porous materials or membranes to separate components based on size, shape, or charge. These techniques exploit differences in molecular size and
permeability to achieve separation.

Answer 7

Filtration

Question 8

(HIC) stands for

Answer 8

Hydrophobic Interaction Chromatography

Question 9

SEC stands for

Answer 9

Size Exclusion Chromatography

Question 10

PAGE stands for

Answer 10

Polyacrylamide Gel Electrophoresis

Question 11

involves determining the concentration of the isolated and purified protein.

Answer 11

PROTEIN QUANTIFICATION

Question 12

2 TECHNIQUES in PROTEIN QUANTIFICATION

Answer 12

1. UV ABSORBANCE
   2.COLOMETRIC ASSAYS

Question 13

measurement at 280 nm relies on the strong
absorption of aromatic amino acids, particularly tyrosine and tryptophan, which are commonly found in proteins.

Answer 13

UV absorbance

Question 14

rely on specific chemical reactions that produce color changes proportional to the protein concentration.

Answer 14

COLOMETRIC ASSAYS

Question 15

Involves the reaction of peptide bonds with copper ions in an alkaline solution.

Answer 15

Biuret Assay

Question 16

Utilizes the binding of Coomassie Brilliant Blue dye to proteins

Answer 16

Bradford Assay

Question 16

It is based on the reduction of Folin-Ciocalteu reagent by protein-bound tyrosine and tryptophan residues, resulting in a blue color complex.

Answer 16

Lowry Assay

Question 17

Relies on the reduction of Cu2+ to Cu1+ by peptide bonds in the presence of bicinchoninic acid, leading to the formation of a purple-colored complex.

Answer 17

BCA Assay