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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
Dyes for Albumin measurement
Methyl orange HABA (2,4-Hydroxybenzone benzoic acid) Bromcresol green Bromcresol purple
26
nonspecific dye for albumin
Methyl orange
27
albumin dye with many interferences (salicylates, bilirubin)
HABA (2,4-Hydroxybenzone benzoic acid)
28
Sensitive dye for albumin Overestimates low alb lvl
Bromcresol Green
29
Most commonly used dye for Albumin
Bromcresol GREEN
30
Specific, sensitive, and precise dye for Albumin
Bromcresol Purple
31
inc. negativity inc. number (conc.) = ?
Inc. migration distance
32
most anodic protein
albumin
33
least anodic protein
gamma globulin
34
support media for protein electrophoresis
Cellulose acetate Agarose gel
35
Inc. conc. of fractionated protein = ?
Darker band fraction
36
Used to compute the area under the absorbance where the medium is placed after electrophoresis
Scanning densitometer
37
Normal: Alb, a1, a2, B INCREASED: y
monoclonal gammopathy
38
Decreased: Alb, a1, B, y Increased: a2
Nephrotic syndrome
39
Normal: B, y Decreased: Alb Increased: a1, a2
Acute inflammation
40
Normal: Alb, a1, a2, B Severely decreased/none: y
Hypogamma globulinemia
41
Normal: Alb, a2, B, y Severely decreased: a1
a1 antitrypsin deficiency
42
Fused increase of B and y all are decreased
Cirrhosis (B-y bridging) *diagnostic
43
Normal: Albumin Increased: a1, a2, B, y
Chronic inflammation
44
Demonstrate other specific proteins IN THE GLOBULIN FRACTION, outside albumin fraction
High resolution protein electrophoreis
45
uses higher voltage couple with a cooling system and more concentrated buffer
High resolution protein electrophoreis
46
Turbidimetric methods for proteins
SSA TCA Benzethonium chloride
47
Proteins are precipitated as particles and turbidity measured spectrophotometrically
Turbidimetric methods: SSA TCA Benzethonium chloride
48
Rapid test easy to use and perform
Turbidimetric methods: SSA TCA Benzethonium chloride
49
Has unequal sensitivity for individual protein
Turbidimetric methods Dye binding
50
Accurate method for urine protein
Biuret
51
Initial biuret reaction
Folin Lowry
52
Oxidation of AA residues by a reagent with a blue resulting color
Folin Lowry
53
Reagent used in Folin Lowry
Folin Phenol reagent
54
Resulting color in Folin Lowry method
blue
55
Very sensitive test for urine protein
Folin Lowry
56
Has limited linearity of the reaction
Dye binding for proteins in other body fluids (Coomassie blue, Ponceau S)
57
diagnostic method for MS
CSF protein electrophoresis
58
diagnostic result for MS
Oligoclonal banding in CSF protein electrophoresis
59
conditions in the blood and tissues outside the CNS
Multiple myeloma (serum/urine) Monoclonal and Polyclonal gammopathies
60
Ave nitrogen content in proteins
16%

MTAP 2 (23 decks)

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