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 = 4 x 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