1. Determination of Serum Total Protein Concentration
224 PHL
Lab#2

2. Classification of Proteins
1) Simple Proteins:
They yield only α-amino acids on hydrolysis.
e.g. albumin, globulin.
2) Conjugated Proteins:
They are proteins combined with a non-protein substance called prosthetic group.
e.g. nucleoprotein, phosphoprotein, metalloprotein, glycoprotein, lipoprotein etc.
3) Derived Protein:
They are derived from changes in original proteins, include denatured proteins.
Plasma Proteins :
Contain both simple and conjugated proteins.
i.e. albumin, globulin, lipoproteins, glycoproteins, metalloproteins.

3. Difference between albumin & globulin
Low molecular weight
High molecular weight (complex molecule)
Soluble in half saturated solution of (NH4)2SO4
Insoluble
2.5 gm/dl
4 gm/dl

4. Serum Total Protein can be determined by two methods
1- Kit method.
2- Biuret method (manual ).

5. Clinical significance:
Hyperproteinemia is high level of serum proteins
Hypoproteinemia is Low level of serum proteins
Causes:
1. Severe hemorrhage.
2. Malnutrition.
3. Malabsorption.
4. High fever causing increased catabolism of proteins.
5. Acute or chronic hepatic insufficiency.
Causes: 1. Dehydration. 2. Chronic inflammations e.g.: tuberculosis 3. Cancer. 4. Drugs e.g.: cortisone, oral contraceptives.

6. Determination of Serum Total Protein Concentration (Kit method)
Principle:
Protein in the sample reacts with the (cu+2) ions in alkaline medium forming a coloured complex that can be measured spectrophotometry.

7. Procedure: Blank = 0.2 ml D.W + 1 ml Reagent-1
Sample = 0.2 ml Sample + 1 ml Reagent-1
Standard = 0.2 ml Standard+1 ml Reagent-1
Mix & stand for 30 min at R.T.
Read the Absorbance for Sample & Standard against the blank at 546 nm.

8. Calculation:
Serum total protein (g/dl)=
Serum Urea conc. = A sample/A standard x Conc. Of St. (58.48 g/l)
Normal value:
(64-83 g/l)