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CC LAB > Proteins_ SERUM PROTEIN ELECTROPHORESIS > Flashcards

Proteins_ SERUM PROTEIN ELECTROPHORESIS Flashcards

1
Q

SERUM PROTEIN ELECTROPHORESIS
• Serum are applied close to the_____ end of a support medium (ph_____)

A

cathode (pH 8.6)

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2
Q

• All major serum proteins carry a net____ charge at ph 8.6 and migrate toward the____.

A

negative

anode

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3
Q

SERUM PROTEIN ELECTROPHORESIS

5 bands:

A

Albumin
Alpha-1 globulins
Alpha-2 globulins
Beta-globulins
Gamma-globulins

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4
Q

: travels farthest to the anode

A

Albumin

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5
Q

= number of proteins present in that fraction.

A

Width of the band

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6
Q

SERUM PROTEIN ELECTROPHORESIS
• After separation, protein fractions are_____ by immersing the support medium in an______

• Next step, proteins are_____.

• Cleared transparent medium is placed in a______ for reading.

A

fixed - acid solution (acetic acid)

stained

scanning densitometer

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7
Q

Dyes used in serum protein electrophoresis

A

Ponceau S
Amido black
Coomassie blue

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8
Q

• The pattern on the membrane moves past a slit through which light is transmitted to a phototube to record the absorbance of the dye that is bound to each protein fraction.

A

scanning densitometer

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9
Q

• Absorbance is recorded on a strip-chart recorder to obtain a pattern of the fraction.

• Compute the area under the absorbance curve for each band and the percentage of total dye that appears in each fraction.

• Concentration is calculated as a percentage of the total protein

• Computation also be made by cutting out the small bands from the membrane and eluting the dye in_____

• Absorbances are added to obtain total absorbance, and the percentage of the total absorbance is calculated

A

0.1 mol/L NaOH

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10
Q

Albumin

Percentage and Concentration

A

53 - 65%

3.5 - 5.0 g/dL

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11
Q

a-1 globulin

Percentage and Concentration

A

2.5 - 5%

0.1 - 0.3 g/dL

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12
Q

a-2 globulins

Percentage and Concentration

A

7 - 13%

0.6 - 1.0 g/dL

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13
Q

Beta-globulin

Percentage and Concentration

A

8 - 14%

0.7 - 1.1 g/dL

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14
Q

Beta-globulin

Percentage and Concentration

A

8 - 14%

0.7 - 1.1 g/dL

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15
Q

Gamma globulin

Percentage and Concentration

A

12 - 22%

0.8 - 1.6 g/dL

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16
Q

” gamma spike”

A

MONOCLONAL IMMUNOGLOBULIN DISEASE

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17
Q

MONOCLONAL IMMUNOGLOBULIN DISEASE

A

Gamma spike

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18
Q

_________ increase in the gamma area, that means there is a_____ so a spike in the Gamma or beta or sometimes in the Alpha 2 region signals the need for examination of immunoglobulins and observed for clinical signs of myelomatosis.

A

monoclonal increase

monoclonal immunoglobulin disease

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19
Q

• Is a condition that raises your risk for lung and other diseases.

• The______ is a protein made in your liver to help protect the lungs so if your body does not make enough Alpha One antitrypsin, your lungs are more easily damaged from smoking pollution or dust.

A

ALPHA-1-ANTITRYPSIN DEFICIENCY

alpha one antitrypsin

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20
Q

• the patient loses serum albumin and low molecular weight proteins in the urine and some IGG
• At the same time an increase occurs in alpha 2 macroglobulin beta lipoprotein components and the haptoglobin
• These events leads to dramatic decrease in the relative amount of albumin and a significant increase in the relative amount of alpha 2 globulin and beta globulin fraction

A

Nephrotic syndrome

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21
Q

result in decrease in albumin and increase in a-1 globulin, a-2 globulin and beta globulin.

A

Inflammation

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22
Q

Also called an Acute Phase Reactant pattern

• These patterns are seen in trauma, burns, infarction, malignancy and liver diseases.

A

Inflammation
| ^
V Albumin | Globulins

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23
Q

• In infectious _____ the gamma globulin fraction rises with increasing hepatocellular damage

• In obstructive jaundice there is an increase in the a-2 and beta-2 globulins. Also noted in the obstructive jaundice is an increased concentration of lipoprotein which is an indicator of its biliary origin.

A

CIRRHOSIS

24
Q

TC N, ↓

A ↓

G ↑

A

Hepatic damage

Burns, trauma

Infections

25
Q

B-y bridging

A

Cirrhosis

26
Q

• ↑ y-globulins

A

Hepatitis

27
Q

• ↑ a2, B-globulins

A

Obstructive jaundice

28
Q

Infections
• ↑ a1-a2-globulins
• ↑ a1-a2-y-globulins

A

Acute

Chronic

29
Q

TC ↓

A ↓

G N

A

Malabsorption
Inadequate diet
Nephrotic syndrome ↑a1-, B-globulins; ↓y-globulins

30
Q

TC ↓

A N

G ↓

A

Immunodeficiency syndromes

31
Q

TC ↓

A ↓

G ↓

A

Salt retention svndrome

32
Q

TC ↑

A ↑

G ↑

A

Dehydration

33
Q

TC ↑

A N

G ↑

A

Multiple myeloma
Monoclonal and polyclonal gammopathies

34
Q

• Modifying electrophoretic parameters
(12 bands)

• Uses higher voltage coupled with a cooling system and more conc. buffer.

A

HIGH-RESOLUTION PROTEIN
ELECTROPHORESIS

35
Q

HIGH-RESOLUTION PROTEIN
ELECTROPHORESIS

•_______- support medium
•_____ estimates of protein
• useful in detecting____ monoclonal bands and differentiating unusual bands

A

Agarose gel

semiquantitative

small

36
Q

• Separation of molecules takes place in silica capillaries
• Capillaries are typically 30-50 cm long

A

CAPILLARY ELECTROPHORESIS

37
Q

CAPILLARY ELECTROPHORESIS

• Separation of molecules takes place in____
• Capillaries are typically____ long

A

silica capillaries

30-50 cm long

38
Q

• Zone electrophoresis that separates proteins on the basis of pl

A

ISOELECTRIC FOCUSING

39
Q

• Specific proteins may be identified by immunochemical assays in which the reaction of protein (antigen) and antibody is measured.

A

IMMUNOCHEMICAL METHODS

40
Q

IMMUNOCHEMICAL METHODS (6)

A

Radial immunidiffusion

immunoelectrophoresis

immunofixation electrophoresis

electroimmunodiffusion

immunoturbidimetry

immunonephelometry

41
Q

IMMUNOCHEMICAL METHODS

Fluorophore
Enzyme
Gold particles

A

Marker

42
Q

IMMUNOCHEMICAL METHODS

Antigen

A

Target protein

43
Q

Inside a damaged kidney there is a presence of

A

Albumin

44
Q

• resulting from glomerular or tubular dysfunction

A

Proteinuria

45
Q

URINARY PROTEINS

Qualitative test:
Quantitative test:

A

Reagent test strip

12-24 hour urine specimen

46
Q

URINARY PROTEINS

Precipitation methods:

A

Sulfosalicylic acid (SSA)
Trichloroacetic acid (TCA)
Benzethonium chloride

47
Q

URINARY PROTEINS

Chemical Methods:

A

Biuret Method
Folin-Lowry Method

48
Q

URINARY PROTEINS

Dye-binding methods:

A

Coomasie blue
Ponceau S

49
Q

Proteins are precipitated as fine particles, turbidity is measured spectrophotometrically

Rapid, easy to use; unequal sensitivity for individual protein

A

Turbidimetric methods (sulfosali-cylic acid, trichloroacetic acid, or benzethonium chloride)

50
Q

Proteins are concentrated by precipitation, redissolved in alkali, then reacted with Cu?t; Cu?+ forms colored complex with peptide bonds

Accurate

A

Biuret

51
Q

Initial biuret reaction; oxidation of tyrosine, tryptophan, and histidine residues by Folin phenol reagent (mixture of phosphotungstic and phosphomolybdic acids): measurement of resultant blue color

Very sensitive

A

Folin-Lowry

52
Q

Protein binds to dye, causes shift in absorption maximum

Limited linearity; unequal sensitivity for individual proteins

A

Dye binding (Coomassie blue,
Ponceau S)

53
Q

CSF PROTEINS
Reference interval (10-40 years old):

A

15-45 mg/dL

54
Q

• Abnormal increased of total CSF Proteins:

A

Bacterial, Viral, and Fungal Meningitis
traumatic tap;

multiple sclerosis;

obstruction;

neoplasm;

disk herniation;

cerebral infraction

55
Q

Degree of permeability can be evaluated by measuring the
CSF albumin comparing it with the serum albumin.
(Ref. value CSF albumin-serum albumin ratio: less than_______)

A

2.7 to 7.3

56
Q

CSF PROTEINS

Increased:

Decreased:

A

damaged blood brain barrier

hyperthyroidism, fluid leakage from CNS

CC LAB (46 decks)

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