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MTAP 2 > Protein methods > Flashcards

Protein methods Flashcards

1
Q

Digestion of proteins to measure nitrogen

A

Kjeldahl

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2
Q

TP reference method

A

Kjeldahl

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3
Q

2 steps in Kjeldahl

A

Kjeldahlization
Ammonia measurement (colorimetric)

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4
Q

conversion of N to NH3

A

Kjeldahlization

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5
Q

agents used in Kjeldahlization

A

Sulfuric acid
Cupric sulfate
Potassium sulfate

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6
Q

digesting agent (340-360C)
*kjeldahlization

A

Sulfuric acid

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7
Q

catalyst
*kjeldahlization

A

Cupric sulfate

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8
Q

increase boiling pt
*kjeldahlization

A

Potassium sulfate

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9
Q

2 reactions that may be used in ammonia measurement
*kjeldahl

A

Nessler’s reaction
Berthelot reaction

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10
Q

use of double iodide of Hg and K

A

Nessler’s reaction

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11
Q

Nessler’s reaction

A

Ammonia + Nessler’s rgt –> Ammonium dimercuric iodide (YELLOW)

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12
Q

Berthelot reaction

A

Ammonia + Alk. Hypochlorite –> Indophenol BLUE

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13
Q

measurement of RI

A

refractometry

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14
Q

formation of violet-colored chelate betw. cupric ions and peptide bonds (measured at 540 nm)

A

Biuret

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15
Q

Biuret reagent composition

A

Cupric ions
Tartrate salt
Potassium iodide

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16
Q

Biuret rgt:
Breaks peptide bonds

A

Cupric ions

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17
Q

Biuret rgt:
Keeps copper in soln

A

Tatrate salt

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18
Q

Biuret rgt:
Stabilizes cupric ions

A

Potassium iodide

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19
Q

Protein binds to dye and causes spectral shift in the absorbance max of dye

A

Dye binding

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20
Q

Used dyes for Total protein measurement

A

Bromphenol blue
Coomassie brilliant blue
Lissamine green
Amido black 10B
Ponceau S

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21
Q

Methods for Albumin measurement

A

Salt precipitation
Dye binding
Electrophoresis

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22
Q

precipitating agent in Salt Precipitation

A

Sodium sulfate (precipitates Globulin)

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23
Q

precipitated in high salt conc.

A

Globulin

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24
Q

left in supernatant which will be measured using Biuret reaction

A

Albumin

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25
Q

Dyes for Albumin measurement

A

Methyl orange
HABA (2,4-Hydroxybenzone benzoic acid)
Bromcresol green
Bromcresol purple

26
Q

nonspecific dye for albumin

A

Methyl orange

27
Q

albumin dye with many interferences (salicylates, bilirubin)

A

HABA (2,4-Hydroxybenzone benzoic acid)

28
Q

Sensitive dye for albumin
Overestimates low alb lvl

A

Bromcresol Green

29
Q

Most commonly used dye for Albumin

A

Bromcresol GREEN

30
Q

Specific, sensitive, and precise dye for Albumin

A

Bromcresol Purple

31
Q

inc. negativity inc. number (conc.) = ?

A

Inc. migration distance

32
Q

most anodic protein

33
Q

least anodic protein

A

gamma globulin

34
Q

support media for protein electrophoresis

A

Cellulose acetate
Agarose gel

35
Q

Inc. conc. of fractionated protein = ?

A

Darker band fraction

36
Q

Used to compute the area under the absorbance where the medium is placed after electrophoresis

A

Scanning densitometer

37
Q

Normal: Alb, a1, a2, B
INCREASED: y

A

monoclonal gammopathy

38
Q

Decreased: Alb, a1, B, y
Increased: a2

A

Nephrotic syndrome

39
Q

Normal: B, y
Decreased: Alb
Increased: a1, a2

A

Acute inflammation

40
Q

Normal: Alb, a1, a2, B
Severely decreased/none: y

A

Hypogamma globulinemia

41
Q

Normal: Alb, a2, B, y
Severely decreased: a1

A

a1 antitrypsin deficiency

42
Q

Fused increase of B and y
all are decreased

A

Cirrhosis (B-y bridging)
*diagnostic

43
Q

Normal: Albumin
Increased: a1, a2, B, y

A

Chronic inflammation

44
Q

Demonstrate other specific proteins IN THE GLOBULIN FRACTION, outside albumin fraction

A

High resolution protein electrophoreis

45
Q

uses higher voltage couple with a cooling system and more concentrated buffer

A

High resolution protein electrophoreis

46
Q

Turbidimetric methods for proteins

A

SSA
TCA
Benzethonium chloride

47
Q

Proteins are precipitated as particles and turbidity measured spectrophotometrically

A

Turbidimetric methods:
SSA
TCA
Benzethonium chloride

48
Q

Rapid test easy to use and perform

A

Turbidimetric methods:
SSA
TCA
Benzethonium chloride

49
Q

Has unequal sensitivity for individual protein

A

Turbidimetric methods
Dye binding

50
Q

Accurate method for urine protein

51
Q

Initial biuret reaction

A

Folin Lowry

52
Q

Oxidation of AA residues by a reagent with a blue resulting color

A

Folin Lowry

53
Q

Reagent used in Folin Lowry

A

Folin Phenol reagent

54
Q

Resulting color in Folin Lowry method

55
Q

Very sensitive test for urine protein

A

Folin Lowry

56
Q

Has limited linearity of the reaction

A

Dye binding for proteins in other body fluids
(Coomassie blue, Ponceau S)

57
Q

diagnostic method for MS

A

CSF protein electrophoresis

58
Q

diagnostic result for MS

A

Oligoclonal banding in CSF protein electrophoresis

59
Q

conditions in the blood and tissues outside the CNS

A

Multiple myeloma (serum/urine)
Monoclonal and Polyclonal gammopathies

60
Q

Ave nitrogen content in proteins

MTAP 2 (23 decks)

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