LABORATORY METHOD (TOPIC 2 PROTEINS)

Question 1

Total Protein preferred Specimen

Answer 1

Serum, non-fasting

Question 2

Interferences of Total protein specimen consideration

Answer 2

Lipemia
Hemolysis

Question 3

Low total protein at?

Answer 3

Birth
During pregnancy
Elderly

Question 4

Concentration is slightly low in recumbent position than ambulatory due to shift in water distribution.

Answer 4

Total protein

Question 5

Specimen that can also be used for Total Protein Determination

Answer 5

Urine 24H
CSF

Question 6

Total protein determination Method

Answer 6

Kjeldahl method
Biuret reaction
Lowry method
Dye binding by BRADFORD
Refractometry
Salt precipitation Test
UV absorption
Serum Electrophoresis

Question 7

What TOTAL PROTEIN METHOD
Measure nitrogen content after protein digestion.

Boric ammonia is TITRATED with HCL to determine the amount of nitrogen present in the solution.

Precise and accurate but time consuming.

Answer 7

Kjeldahl method

Question 8

Serum protein is treated with CuSO4 in alkaline medium forming purple/violet color.

color intensity is directly proportional to the concentration of protein in solution.

Answer 8

BIURET REACTION (colorimetric)

Question 9

Most widely used method for TP determination.

Answer 9

BIURET REACTION (colorimetric)

Question 10

Buret Reagent for Biuret Reaction

Answer 10

CuSO4
NaOh
Rochelle Salt (potassium-sodium-tartate)
Kl

Question 11

Biuret Reaction Modifications: Kingsley and Weischelbaum – uses _____

Answer 11

22-26% Na2SO4

Question 12

Lowry Method aka

Answer 12

Folin Ciocalteau

Question 13

based on the reaction of TRYPTOPHAN and TYROSINE residues of protein with Folin ciocalteau amixture of PHOSPHOTUNGTIC ACID and PHOSPOMOLYBDIC ACID producing BLUE colored complex.

Answer 13

LOWRY METHOD (Folin ciocalteau)

Question 14

dye binds amine group of amino acid residues in polypeptide chain

Answer 14

DYE BINDING by BRADFORD

Question 15

Dyes used in DYE BINDING by BRADFORD

Answer 15

Bromcresol purple(BCP)- the most sensitive
Bromcresol green (BCG)- the commonly used
Tetrabromphenol blue- used in urine reagent strip. Specific to albumin
Ninhydrin- specific for amino acids
Pyrogallol red- for urine and CSF protein

Question 16

the most sensitive DYE BINDING by BRADFORD

Answer 16

Bromcresol purple(BCP)-

Question 17

the commonly used DYE BINDING by BRADFORD

Answer 17

Bromcresol green (BCG)

Question 18

used in urine reagent strip. Specific to albumin by DYE BINDING by BRADFORD

Answer 18

Tetrabromphenol blue-

Question 19

specific for amino acids by DYE BINDING by BRADFORD

Answer 19

Ninhydrin

Question 20

for urine and CSF protein by DYE BINDING by BRADFORD

Answer 20

Pyrogallol red-

Question 21

Rapid estimation of protein thru measurement of refractive index.
Used in urine protein determination.

Answer 21

REFRACTOMETRY

Question 22

SALT PRECIPITATION TEST
For urine and CSF protein

1. precipitates all protein
2. precipitates all globulin

Answer 22

Sulfosalicylic acid test (SSA)-
Trichloroacetic acid test (TCA)-

Question 23

based on the protein’s ability to absorb UV light at 280 nm

Answer 23

UV ABSORPTION

Question 24

Normal value of Total Protein

Answer 24

6.5-8.5 g/dl
To convert in g/L multiply by 10

Question 25

most important plasma proteins with low molecular weight compared with most of the other proteins.

Answer 25

Albumin

Question 26

Maintain colloid osmotic pressure of plasma. Transport of amino acids from liver to tissues. Serves as transport protein or carrier protein.

Answer 26

Albumin

Question 27

Most widely used laboratory method for Albumin

Answer 27

Dye binding methods

Question 28

Albumin Normal Value

Answer 28

3.5-5.5 g/dl

Question 29

Clinical significance absence of albumin

Answer 29

Analbuminemia

Question 30

presence of 2 bands in the albumin region (electrophoresis)

Answer 30

Bisalbuminemia

Question 31

decrease level of albumin

Answer 31

Hypoalbuminemia

Question 32

Clnical Sinificance Albumin increase in?

Answer 32

Shock Acute dehydration Multiple Sclerosis Hypothyroidism Addisons disease Multiple myeloma Diarrhea

Question 33

Clnical Sinificance Abumin Decrease in?

Answer 33

Malnutrition and Mal-absorption Liver disease Nephrotic syndrome Burns Extensive bleeding Edema Last two trimester of pregnancy

Question 34

Group of 20 or more proteins grouped on the basis of solubility

Answer 34

Globulins

Question 35

Globulins consists of?

Answer 35

Alpha and Beta globulins Gamma globulins

Question 36

Globulin synthesized in the liver

Answer 36

Alpha and Beta globulin

Question 37

Globulin synthesized in the plasma cells

Answer 37

Gamma globulin

Question 38

Globulins determination separate ALBUMIN and GLOBULIN globulins precipitate in 27% NaSO4 while albumin stays in solution

Answer 38

DIFFERENTIAL SOLUBILITY OF SALT SOLUTION

Question 39

Globulins determination direct colorimetry TRYPTOPHAN residue reacts with GLYOXYLIC ACID in the presence of STRONG ACID MEDIUM producing purple color and read at 555 nm

Answer 39

HOPKINS-COLE METHOD

Question 40

Globulins determination Determine the concentration of TOTAL PROTEIN and ALBUMIN GLOBULIN is determined indirectly by difference.

Answer 40

BIURET METHOD

Question 41

Globulin Biuret method Normal Value

Answer 41

2.0-3.5 g/dl

Question 42

Globulin Formula

Answer 42

Total protein - albumin

Question 43

use to measures the total amount of plasma proteins

Answer 43

A/G RATIO (ALBUMIN/GLOBULIN)

Question 44

A/G RATIO (ALBUMIN/GLOBULIN) normal value

Answer 44

1.1-1.8:1

Question 45

A/G ratio increase in?

Answer 45

Dehydration High protein diet Corticosteroid Growth Hormone

Question 46

A/G ratio decrease in?

Answer 46

Liver disease Infectious diseases Multiple myeloma Nephrotic syndrome