Total Protein Concentration 224 PHL Lab 2 1 Simple Proteins They yield only α amino acids on hydrolysis eg albumin globulin 2 Conjugated Proteins They are proteins combined with a nonprotein substance called prosthetic group ID: 502989
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Slide1
Determination of Serum Total Protein Concentration
224 PHLLab#2 Slide2
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.
Classification of ProteinsSlide3
Albumin
GlobulinLow molecular weight
High molecular weight (complex molecule)Soluble in half saturated solution of
(NH4)
2
SO4Insoluble2.5 gm/dl4 gm/dlDifference between albumin & globulinSlide4
1- Kit method. 2- Biuret method (manual ).
Serum Total Protein can be determined by two methodsSlide5
Clinical significance:
Hyperproteinemia is high level of serum proteins
Hypoproteinemia is
Low level of serum proteins
Causes
: 1. Dehydration. 2. Chronic inflammations e.g.: tuberculosis3. Cancer.4. Drugs e.g.: cortisone, oral contraceptives.Causes: 1. Severe hemorrhage.2. Malnutrition.3. Malabsorption
.
4. High fever causing increased catabolism of proteins.
5. Acute or chronic
hepatic insufficiency
.Slide6
Principle:Protein in the sample reacts with the
(cu+2) ions in alkaline medium forming a coloured complex that can be measured spectrophotometry.
Determination of
Serum Total Protein Concentration(Kit method)Slide7
Blank = 0.2 ml D.W + 1 ml Reagent-1
Sample = 0.2 ml Sample + 1 ml Reagent-1Standard =
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.Procedure:Slide8
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)