Amino Acids & Plasma Proteins Flashcards by Enrico Aque

Total Protein sample of choice

Serum

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Classic method for Total Protein

Kjeldahl Method

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Most widely used method for Total Protein

Biuret method

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IFCC stands for

International Federation of Clinical Chemistry

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In (TP) biuret method, what colored chelate is formed

violet

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(TP) Biuret reagent

Potassium hydroxide

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what color is resulted from the reaction of the biuret method for total protein

Pink to reddish violet

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In (TP) Dye Binding Method, Absorbance at ______ is used to determine the protein concentration

590nm

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A quick alternative to chemical analysis of serum total protein where a rapid estimate is required

Refractometry

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4 Total Protein Methods

Kjeldahl
Biuret
Dye Binding
Refractometry

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Which Total Protein Method:

Digestion of protein; Measurement of nitrogen content

Kjeldahl

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Which Total Protein Method:

Formulation of violet-colored chelate between Cu2+ ions and peptide bonds

Biuret

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Total Protein - Albumin = ?

Globulin

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Decrease in A/G ratio may be resulted by

Decreased Albumin or Increased Globulin

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Elevated A/G ratio may be resulted by

Increased Albumin or Decreased Globulin

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Globulins are separated from albumin by salting out, using sodium salt to precipitate globulins

Salt Fractionation

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Most widely used method for determining albumin

Dye Binding

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Absorbance of the albumin-dye complex is equal to?

Amount of Albumin

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4 dyes used in Dye Binding for Measuring Protein Factors

Methyl Orange
2,4’ - hydroxyazobenzene-benzoic acid
Bromocresol Green
Bromocresol Purple

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Dye used in Dye Binding for Measuring Protein Factors:

Not specific for albumin

Methyl Orange

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Dye used in Dye Binding for Measuring Protein Factors:

Low sensitivity but more specific for albumin

2,4’-hydroxyazobenzene-benzoic acid

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Dye used in Dye Binding for Measuring Protein Factors:

Not affected by interfering substances
Hemoglobin binds to it which can lead to falsely high albumin values

Bromocresol Green

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Dye used in Dye Binding for Measuring Protein Factors:

Binds specifically to albumin

Bromoscresol Purple

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Total Protein - Globulin = ?

Albumin

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In Total Globulins, Glyoxylic acid condenses with tryptophan to produce what color

Purple

For when an abnormality in the total protein or albumin concentration is found

Electrophoresis

Electrophoresis: Protein charge for when pH is greater than the isoelectric point

negative

Electrophoresis: Protein charge for when pH is less than the isoelectric point

positive

The Anion protein will migrate to the?

Anode

The Cation protein will migrate to the?

Cathode

Electrophoresis: What can be estimated from the difference between the isoelectric point of the protein and the pH of the buffer?

Speed of Migration

In the electrophoretic mobility formula, what does "s" stand for

Distance traveled in centimeters

In the electrophoretic mobility formula, what does "t" stand for

Time of migration in seconds

In the electrophoretic mobility formula, what does "F" stand for

Field strength in velocity over centimeters

Before, electrophoresis used an aqueous medium and was called?

Moving boundary electrophoresis

Electrophoresis: Supporting media in today's clinical laboratories

Cellulose Acetate or Agarose Gel

Electrophoresis: Paper was used as a solid medium in a process known as?

Zone Electrophoresis

4 methods for measuring protein factors

Salt Fractionation Dye Binding Total Globulin Electrophoresis

All major serum proteins carry a net charge at what pH

pH 8.6

5 bands of Serum Protein Electrophoresis

Albumin alpha 1-Globulin alpha 2-Globulin beta-Globulin gamma-Globulin

Serum Protein Electrophoresis: band the travels farthest to the anode

Albumin

Serum Protein Electrophoresis: The number of proteins present in that fraction can be determined by the

Width on the band

Dyes used in Serum Protein Electrophoresis

Ponceau S Amido black Coomassie Blue

Serum Protein Electrophoresis: Reference percentage for Albumin

53% - 65%

Serum Protein Electrophoresis: Reference percentage for a1-globulin

2.5% - 5%

Serum Protein Electrophoresis: Reference percentage for a2-globulin

7% - 13%

Serum Protein Electrophoresis: Reference percentage for b-globulin

8% - 14%

Serum Protein Electrophoresis: Reference percentage for g-globulin

12% - 22%

Serum Protein Electrophoresis: Reference concentration for Albumin

3.5 - 5.0 g/dL

Serum Protein Electrophoresis: Reference concentration for a1-globulin

0.1 - 0.3 g/dL

Serum Protein Electrophoresis: Reference concentration for a2-globulin

0.6 - 1.0 g/dL

Serum Protein Electrophoresis: Reference concentration for b-globulin

0.7 - 1.1 g/dL

Serum Protein Electrophoresis: Reference concentration for g-globulin

0.8 - 1.6 g/dL

Densitometric pattern that is indicates Monoclonal Immunoglobulin Disease

Monoclonal Increase

Monoclonal Increase is aka

Gamma Spike

Protein made in your liver to help protect the lungs

alpha-1-antitrypsin

Inflammation densitometric pattern is also called an?

Acute Phase Reactant pattern

Densitometric pattern: ↑ g-globulin

Monoclonal Increase

Densitometric pattern: ↓ a1-gloublin

Alpha-1-Antitrypsin Deficiency

Densitometric pattern: ↓ Albumin ↑ a2-globulin ↑ b-globulin

Nephrotic Syndrome

Densitometric pattern: ↓ Albumin ↑ a1-gloublin ↑ a2-globulin ↑ b-globulin

Inflammation

Densitometric pattern: ↓ Albumin beta-gamma bridge ↑↑ g-globulin

Cirrhosis

High-Resolution Protein Electrophoresis has how many bands

Collection of technique in which the separation of molecules takes place in silica capillaries

Capillary Electrophoresis

Support Medium commonly used in High-Resolution Protein Electrophoresis

Agarose Gel

Zone electrophoresis that separates proteins on the bases of isoelectric point

Isoelectric Focusing

Isoelectric focusing used what medium

Polycrylamide or Agarose Gel

Plasma proteins originating from the kidney and urinary tract or from other sources such as the vagina and prostate

Urinary Protein

Urinary Protein (color): 1. Absence of Protein = 2. Increased concentration of protein =

1. Yellow 2. Blue

Reference interval for CSF Protein for 10-40yrs old

14-45mg/dl

Ref value for CSF albumin-serum albumin ratio

Less than 2.7 to 7.3

Increased CSF albumin-serum albumin ratio

Damaged blood brain barrier

Decreased CSF albumin-serum albumin ratio

Hyperthyroidism

Incubator temperature

37 degrees Celsius

Incubation time for Total Protein

10mins

Incubation time for Total Albumin Ratio

1min

Incubation time for Glucose Procdure

10min

Amino Acids & Plasma Proteins Flashcards

(77 cards)