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TOTAL PROTEIN Definition of protein:- A large group of nitrogenous organic compounds that are essential constituents of living cells; consist of polymers.

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Presentation on theme: "TOTAL PROTEIN Definition of protein:- A large group of nitrogenous organic compounds that are essential constituents of living cells; consist of polymers."— Presentation transcript:

1 TOTAL PROTEIN Definition of protein:- A large group of nitrogenous organic compounds that are essential constituents of living cells; consist of polymers of amino acids (are joined together by the peptide bonds); essential in the diet of animals for growth and for repair of tissues; can be obtained from meat, eggs and milk...

2 Total Serum Protein:- A total serum protein test measures the total amount of protein in the blood. It also measures the amounts of two major groups of proteins in the blood: albumin and globulin.  Types:- 1)Albumin. 2)α1globulin (α1 Antitrypsin) 3)α2globulin ( haptoglobulin) 4)β globulin ( transferrin, fibrinogen) 5)γ globulin ( Immunoglobulins).

3 On the basis of functions of protein: 1. Structural protein: e.g. keratin 2. Enzyme and catalytic protein: e.g. pepsin 3. Transport protein: Hb, serum albumin 4. Hormonal protein: e.g. hormones as insulin, adrenalin 5. Contractile protein: e.g. actin and myosin 6. Storage protein: e.g. oval albumin, glutamine 7. Genetic protein: e.g. nucleic acid ( DNA & RNA) 8. Defence protein: e.g. Ig (immunoglobulins). 9. Receptor protein: hormones.

4 Source:- Albumin and most of α and β globulins are formed in liver.. Immunoglobulins are synthesized by the plasma cells in lymph nodes, bone marrow and spleen. Normal values:- total serum protein in normal adult is 6-8 g/dl.

5 In the assay of total serum proteins, useful diagnostic information can be obtained by determining the albumin and the globulin fraction (albumin/globulin ratio).. Serum globulin can be separated into several subgroups by serum protein electrophoresis. Albumin is tested for liver and kidney diseases, globulin is tested for multiple myeloma.

6 Methods for determination of total protein in serum 1.Kjeldahl method. 2.Refractometry. 3.Biuret. 4.Dye Binding. 5.Ultraviolet Absorption.

7 Protein in other body fluids A.Urine protein:  proteinuria: due to concentration of total protein in urine > 12mg/dl. B. CSF protein.

8 Experiment: Total protein( Biuret method) Principle: Cupric ions (Cu+ 2 ) of Biuret reagent will complex with the groups involved in the peptide bond, in an alkaline medium and in the presence of at least 2 peptide bonds, a violet-colored complex is formed. The reagent also contains:  sodium potassium tartrate, to complex (Cu+2) to prevent their precipitation in the alkaline solution.

9  Potassium iodide, which acts as an antioxidant. Procedure: blank standard sample1 sample2 1ml R 1ml R+ 1ml R+ 1ml R+ 20μl std. 20μl S1 20μl S2

10 Mix, stand 10 min. (R.T) Read the absorbance at 540nm. Calculations: {Sample (A)/ Std (A)} * 5= g/dl

11 Clinical significance: 1.Hypoproteinemia or Hemodilution: less than the reference range, occur in : starvation, malabsorption and burns. 2.Hyperproteinmia or Hemoconcentration: Increasing. Occurs due to dehydration ( diarrhea, vomiting.) and excess synthesis of plasma proteins (multiple myeloma)...

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