Total Protein (Biuret method)

 Clinical Significance :-

                         Protein are constitution of muscles enzymes, hormones and several other key functional and structure structural entities in the body . They are involved in the maintenance of the normal distribution of water between blood and tissues. Consisting mainly of albumin and globulin in fractions very independently and widely in diseases. Increased level are found mainly in the  dehydration. Decrease level are found mainly in malnutrition a synthesis,  protein losses as in hemorrhage or excessive protein catabolism.

Principle :-

             Proteins in an alkaline medium, bind with the cupric ions present in the biuret reagent to form a blue blue violet colour Complex. The intensity of the colour formed is directly proportional to the amount of protein present in the sample.

 Material required :-

• Clean and dry glassware.                                     
• Laboratory glass Pipettes or  micro Pipettes and tips. 
• Colorimeter.

 Normal Value :- 

                             6.0 - 8.0 gm/dl   

                         

Sample :-

              Serum , Heparinized/EDTA Blood. 

Procedure :-

                       Pipette into clean and dry test tubes labeled as Blank ( B ) , Standard ( S ), and Test ( T ) :

Addition           Sequence	B	S	T
Reagent	1ml	1ml	1ml
Standard	-	10 ul	-
Sample	-	-	10ul

 Mix well  and  incubate  at  37 ‘ C for 5 mins. Measure absorbance of the standard ( Abs . S ) and Test ( Abs . T ) against Reagent blank.

 

   Calculation  :-

                                              Abs . T

                     Protein ( mg/dl ) =    ..............  * 6

                                              Abs . S   

Preparation and reagent and stability :-

                                        All Reagent are ready to use and are stable till the expiry date mentioned on the label , when stored at 2' to 8' C.

Quality Control :-

                  For accuracy it is necessary to run known controls with every assay.

Limitation & Precautions :-

 

1. Storage conditions as mentioned on the kit to be adhered.
2. Do not freeze or expose the reagent to high temperature and protect from direct as it may affect the performance of the kit.
3. Use clean glassware free from dust .
4. Re-plug the Glucose standard vial after use.

 General System Parameters:-

          Reaction type           : End point 

          Wave length             : 546 nm ( 530 - 570 nm )

          Temperature             : 37'C

          Incubation              : 05 minutes 

          Reagent Volume          : 1.0 ml

          Sample Volume           : 10 ul 

          Standard concentration  : 6 mg/dl

          Zero setting            : Reagent Blank

          Light Path              : 1.0 cm