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    Total Protein Kit

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    Dinesh Sreedharan
    This document provides instructions for determining total protein levels in serum or plasma using a biuret method. It describes that the biuret method involves mixing protein samples with copper sulfate to form a violet colored complex, and the intensity of the color is directly proportional to the total protein concentration. Reagents, samples, standards, assay procedure, calculations, reference values and notes are described to perform the test and analyze results.

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    Total Protein Kit

    Uploaded by

    Dinesh Sreedharan
    75 views2 pages
    This document provides instructions for determining total protein levels in serum or plasma using a biuret method. It describes that the biuret method involves mixing protein samples with copper sulfate to form a violet colored complex, and the intensity of the color is directly proportional to the total protein concentration. Reagents, samples, standards, assay procedure, calculations, reference values and notes are described to perform the test and analyze results.

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    Original Title

    TOTAL PROTEIN KIT

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    This document provides instructions for determining total protein levels in serum or plasma using a biuret method. It describes that the biuret method involves mixing protein samples with copper sulfate to form a violet colored complex, and the intensity of the color is directly proportional to the total protein concentration. Reagents, samples, standards, assay procedure, calculations, reference values and notes are described to perform the test and analyze results.

    Copyright:

    © All Rights Reserved
    Download as DOC, PDF, TXT or read online from Scribd
    Flag for inappropriate content
    Download as doc, pdf, or txt
    75 views2 pages

    Total Protein Kit

    Uploaded by

    Dinesh Sreedharan
    This document provides instructions for determining total protein levels in serum or plasma using a biuret method. It describes that the biuret method involves mixing protein samples with copper sulfate to form a violet colored complex, and the intensity of the color is directly proportional to the total protein concentration. Reagents, samples, standards, assay procedure, calculations, reference values and notes are described to perform the test and analyze results.

    Copyright:

    © All Rights Reserved
    Download as DOC, PDF, TXT or read online from Scribd
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    of 2

    TOTAL PROTEIN

    (Biuret Method)
    For the determination of Total Protein in serum and plasma
    (For In vitro Diagnostic use only)

    SUMMARY:

    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 normal distribution of
    water between blood and the tissues. Consisting mainly of albumin and globulin fractions vary
    independently and widely in diseases.

    PRINCIPLE:

    Protein + Cu++ Blue violet colored complex


    The intensity of the color formed is directly proportional to the amount of total protein present in
    the sample.

    CONTENTS:

    PACK BIURET REAGENT (A1) STANDARD (S)


    SIZE

    2x50ml 2x50ml 2ml

    STORAGE/STABILITY:

    Reagent is stable at R.T,(Below 30˚C) till the expiry mentioned on the label.

    Standard is stable at 2-8˚C till the expiry mentioned on the label.

    REAGENT PREPARATION:

    Reagents are ready to use. Protect from bright light.

    SAMPLE:

    Serum or Plasma. Total Proteins are reported to be stable in the sample for 6 days when stored at
    2-8˚C.
    GENERAL SYSTEM Parameter:

    Reaction Mode End Point Sample Volume 10 µl


    Wavelength 555 nm Reagent volume 1000µl
    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

    ASSAY PROCEDURE:
    Wavelength/ Filter : 555 nm (Hg 546 nm)/Green (520 – 570nm)
    Temperature : R.T/37˚C
    Light Path : 1 cm
    Pipette into clean dry test tube labeled as Blank(B),Standard (S) and Test(T)

    Addition Sequence B(µl) S(µl) T(µl)


    T.Protein Reagent(A1) 1000 1000 1000
    Standard(S) - 10 -
    Sample - - 10
    Mix well and incubate at 37˚C for 5 min or at R.T. for 10 min. Measure the absorbance of the
    standard (Absorbance of Standard) and test Sample (Absorbance of Test) against reagent blank at
    555 nm (520-570 nm) within 60 min.

    CALCULATION:
    Absorbance of Test
    Total Protein in g/dl =------------------------------- × 6
    Absorbance of Standard

    NORMAL REFERENCE VALUES:


    Serum and Plasma : 6.0 – 8.0 g/dl
    It is recommended that each laboratory establish its own normal range representing its patient
    population.

    LINEARITY:
    This procedure is linear up to 15 g/dl. If values exceed this limit, dilute the sample with distilled
    water and repeat assay. Calculate the value using the proper dilution factor.

    NOTE:
    Do not use if the reagent shows turbidity or black precipitates.

    REFERENCES:
    1.Gomali,A.G(1949) Biol.Chem 177:751
    2.Doumas, B.T (1975) Clin Chem 21:1159

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