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    Estimation of Total Protein in Serum &

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    Nada hasan
    This document describes methods for estimating total protein and albumin levels in serum. Total protein is measured using the Biuret method, which detects protein levels colorimetrically based on reaction with copper ions. Albumin is measured using the BCG method, where albumin forms a colored complex with bromocresol green. Both tests involve mixing serum samples with reagents and measuring absorbance to determine protein concentrations, which are important for assessing liver and kidney function and nutritional status. Normal total protein ranges are 6.6-8.7 g/dL for adults and albumin is normally 3.8-5.1 g/dL.

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    Estimation of Total Protein in Serum &

    Uploaded by

    Nada hasan
    904 views22 pages
    This document describes methods for estimating total protein and albumin levels in serum. Total protein is measured using the Biuret method, which detects protein levels colorimetrically based on reaction with copper ions. Albumin is measured using the BCG method, where albumin forms a colored complex with bromocresol green. Both tests involve mixing serum samples with reagents and measuring absorbance to determine protein concentrations, which are important for assessing liver and kidney function and nutritional status. Normal total protein ranges are 6.6-8.7 g/dL for adults and albumin is normally 3.8-5.1 g/dL.

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    This document describes methods for estimating total protein and albumin levels in serum. Total protein is measured using the Biuret method, which detects protein levels colorimetrically based on reaction with copper ions. Albumin is measured using the BCG method, where albumin forms a colored complex with bromocresol green. Both tests involve mixing serum samples with reagents and measuring absorbance to determine protein concentrations, which are important for assessing liver and kidney function and nutritional status. Normal total protein ranges are 6.6-8.7 g/dL for adults and albumin is normally 3.8-5.1 g/dL.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    904 views22 pages

    Estimation of Total Protein in Serum &

    Uploaded by

    Nada hasan
    This document describes methods for estimating total protein and albumin levels in serum. Total protein is measured using the Biuret method, which detects protein levels colorimetrically based on reaction with copper ions. Albumin is measured using the BCG method, where albumin forms a colored complex with bromocresol green. Both tests involve mixing serum samples with reagents and measuring absorbance to determine protein concentrations, which are important for assessing liver and kidney function and nutritional status. Normal total protein ranges are 6.6-8.7 g/dL for adults and albumin is normally 3.8-5.1 g/dL.

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    Download as PPTX, PDF, TXT or read online from Scribd
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    The document discusses methods for estimating total protein and albumin levels in serum or plasma through colorimetric assays. Total protein is comprised of albumin and globulin proteins. Albumin functions to maintain blood pressure and transport medications and nutrients. Globulins include alpha, beta, and gamma types which have roles in binding hemoglobin, fighting infections, and transporting metals.

    The two major groups of proteins measured are albumin and globulin.

    Albumin is mainly produced in the liver and functions to maintain blood pressure by preventing leakage from blood vessels. It also helps transport some medications through the blood and is important for tissue growth and healing.

    Estimation of Total Protein in Serum &

    Estimation of Albumin in Serum


    Estimation of Total Protein in Serum

    This is a photometric colorimetric test for


    .total proteins by Biuret Method
    Introduction:
    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
    Albumin:
    It is made mainly in the liver. It helps
    keep the blood from leaking out of blood
    vessels. Albumin also helps carry some
    medicines through the blood and is
    .important for tissue growth and healing
    Globulin:

    It is made up of different proteins called alpha, beta,


    and gamma types. Some globulins are made by the
    liver, while others are made by the immune system.
    Certain globulins bind with hemoglobin. Other
    globulins transport metals, such as iron, in the blood
    and help fight infection. Serum globulin can be
    separated into several subgroups by serum protein
    .electrophoresis
    Why It Is Done?

    Total protein is tested to:


    1. Check how well the liver and kidney are
    working.
    2. Find out if your diet contains enough
    protein.
    3. Determine your chances of developing an
    infection.
    4. See if you have a rare blood disease,
    such as multiple myeloma.
    Principle:
    Cupric ions react with protein in alkaline
    solution to form a purple complex. The
    absorbance of this complex is proportional
    .to the protein concentration in the sample
    Reagents:
    The reagents and standard provided in
    the kit are ready for use.
    Specimen:
    Serum, heparinized or EDTA plasma can
    .be used
    Assay Parameters:
    Wavelength: Hg 546 nm
    Optical path: 1 cm
    Temperature: 20 - 25°C
    Measurement: Against reagent blank
    Protocol:
    1. Take three cuvettes – label the first one as reagent
    blank, the second as standard and the last one as
    sample.
    2. To the reagent cuvette, add 1000 µl of the reagent
    only.
    3. To the standard cuvette, add 20 µl standard and
    1000 µl reagent.
    4. To the sample cuvette, add 20 µl sample and 1000
    .µl reagent
    5. Mix the three cuvettes and incubate for
    10 min. at 20 - 25°C.
    6. Measure the absorbance of the sample
    and the standard against the reagent
    blank within 30 min.
    7. The measured value will yield the value
    of change in absorbance or ΔA.
    Calculation of Total Protein
    Concentration:
    with Factor:
    C = 19 x ΔA sample [ g/dl ]
    C = 190 x ΔA sample [ g/l ]
    with Standard:
    C = 8 x ΔA sample [ g/dl ]
    ΔA STD

    C = 80 x ΔA sample [ g/l ]
    ΔA STD
    Normal Ranges in Serum or Plasma:
    Infants: 4.6 – 7.0 g/dl or 46 –70 g/l.
    .Adults: 6.6 – 8.7 g/dl or 66 – 87 g/l
    Estimation of Albumin in Serum

    This is a photometric colorimetric test for

    albumin by BCG method


    Principle:
    Bromocresol green forms with albumin in
    citrate buffer a colored complex. The
    absorbance of this complex is
    proportional to the albumin concentration
    .in the sample
    Reagents:
    The reagents and standard provided in the
    kit are ready for use.
    Specimen:
    Serum, heparinized or EDTA plasma can be
    used
    Assay Parameters:
    Wavelength: Hg 546 nm, 578 nm
    Optical path: 1 cm
    Temperature: 20 - 25°C
    Measurement: Against reagent blank
    Protocol:
    1. Take three cuvettes – label the first one
    as reagent blank, the second as standard
    and the last one as sample.
    2. To the reagent cuvette, add 1000 µl of the
    reagent only.
    3. To the standard cuvette, add 10 µl
    .standard and 1000 µl reagent
    4. To the sample cuvette, add 10 µl sample
    and 1000 µl reagent.
    5. Mix the three cuvettes and incubate for 5
    min. at 20 - 25°C.
    6. Measure the absorbance of the sample and
    the standard against the reagent blank
    within 30 min.
    7. The measured value will yield the value of
    change in absorbance or ΔA.
    Calculation of the Albumin
    Concentration:
    C=4x ΔA sample [ g/dl ]
    ΔA STD

    C = 40 x ΔA sample [ g/l ]
    ΔA STD
    Normal Ranges in Serum or Plasma:
    3.8 – 5.1 g/dl or 38 – 51 g/l

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