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Total Protein

This document provides instructions for determining total protein levels in serum or plasma using a biuret method. It describes the principle, reagents, procedure, calculations, reference values and specifications for the assay.

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Dinesh Sreedharan
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64 views1 page

Total Protein

This document provides instructions for determining total protein levels in serum or plasma using a biuret method. It describes the principle, reagents, procedure, calculations, reference values and specifications for the assay.

Uploaded by

Dinesh Sreedharan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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TOTAL PROTEIN ASSAY PROCEDURE:

(Biuret Method)
Wavelength/ Filter : 555 nm (Hg 546 nm)/Green (520 - 570nm)
For the determination of Total Protein in serum and plasma
(For In vitro Diagnostic use only) Temperature : R.T/37°C
Light Path : 1 cm
SUMMARY:
Pipette into clean dry test tube labeled as Blank(B),Standard (S) and Test(T)
Proteins are constituents of muscle, enzymes, hormones and several other key
functional and structural entities in the body. They are involved in the maintenance of the Addition Sequence B(µl) S(µl) T(µl)
normal distribution of water between blood and the tissues. Consisting mainly of albumin T.Protein Reagent(A1) 1000 1000 1000
and globulin fractions vary independently and widely in diseases. Standard(S) - 10 -
Sample - - 10
PRINCIPLE:
Protein + Cu++ Blue violet colored complex Mix well and incubate at 37°C for 5 min or at R.T. for 10 min. Measure the absorbance
The intensity of the color formed is directly proportional to the amount of total protein of the standard (Absorbance of Standard) and test Sample (Absorbance of Test)
present in the sample. against reagent blank at 555 nm (520-570 nm) within 60 min.

CALCULATION:
CONTENTS: Absorbance of Test
PACK SIZE BIURET REAGENT (A1) STANDARD (S) Total Protein in g/dl = ------------------------------------------- × 6
Absorbance of Standard
2x50ml 2x50ml 2ml
STORAGE / STABILITY: NORMAL REFERENCE VALUES:
Reagent is stable at R.T,(Below 30°C) till the expiry mentioned on the label. Serum and Plasma : 6.0 - 8.0 g/dl
Standard is stable at 2-8°C till the expiry mentioned on the label. It is recommended that each laboratory establish its own normal range representing its
patient population.
REAGENT PREPARATION:
Reagents are ready to use. Protect from bright light. LINEARITY:
This procedure is linear up to 15 g/dl. If values exceed this limit, dilute the sample with
SAMPLE: distilled water and repeat assay. Calculate the value using the proper dilution factor.
Serum or Plasma. Total Proteins are reported to be stable in the sample for 6 days when
stored at 2-8°C. NOTE:
Do not use if the reagent shows turbidity or black precipitates.
GENERAL SYSTEM PARAMETER:
REFERENCES:
Reaction Mode End Point Sample Volume 10 µl 1. Gomali,A.G(1949) Biol.Chem 177:751
Wavelength 555 nm Reagent volume 1000µl 2. Doumas, B.T (1975) Clin Chem 21:1159
Blank Reagent Standard 6 g/dl
Incubation 5 min 37°C/10 min R.T Reaction Slope Increasing
Delay Time 5 Sec Linearity 15 g/dl
Read Time 5 Sec Units g/dl

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