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CC1 LECTURE DEAN > PROTEIN DETERMINATION > Flashcards

PROTEIN DETERMINATION Flashcards

1
Q

determination of amino Nitrogen derived from protein

A

KJELDAHL METHOD

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2
Q

Functional groups:

A

amino group and carboxyl group

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3
Q

KJELDAHL METHOD

Nitrogen converted to [?] using [?]

A

NH3 [ammonium]
H2SO4 [bisulfate]

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4
Q

Protein + H2SO4 ——> NH4 + H2SO4

A

KJELDAHL METHOD

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5
Q

(NH4) ——> Na2SO4 + NH3 + 2H2O

A

KJELDAHL METHOD

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6
Q

KJELDAHL METHOD

The nitrogen in ammonia may be measured using:

A

Nesslerization
Berthelot reaction
Titration method

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7
Q
  • basis for measuring the amount of protein] HSO4 + 2NaOH [sodium hydroxide
A

ammonia

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8
Q

ammonia reacts with Nessler’s reagent or double iodide salt of potassium and mercury

A

Nesslerization

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9
Q

Urea and ammonia determination

A

Nesslerization
Berthelot reaction

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10
Q

ammonia reacts with phenol/phenolic cpd and a hypochlorite

A

Berthelot reaction

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11
Q

Berthelot reaction End product:

A

Indophenol

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12
Q

Copper binds to the peptide bond

A

BIURET METHOD

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13
Q

(+) purple-color

A

BIURET METHOD

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14
Q

= purple chromogen

A

Protein + CuSO4 + NaOH + Rochelle salt

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15
Q

Employed spectrophotometric methods

A

BIURET METHOD

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16
Q

Tyrosine & tryptophan reduce PT-PMA reagent = blue color

A

FOLIN-CIOCALTEU METHOD

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17
Q

Detects proteins in low conc. (10-60 pig/ )

A

FOLIN-CIOCALTEU METHOD

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18
Q

Widely used in research to measure tissue & enzyme proteins

A

FOLIN-CIOCALTEU METHOD

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19
Q

Not used in routine laboratory work

A

FOLIN-CIOCALTEU METHOD

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20
Q

Notsatisfactory for urine & CSF protein determination

A

FOLIN-CIOCALTEU METHOD

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21
Q

PT-PMA

A

phosphotungstic phosphomolybdic acid

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22
Q

Dueto the high electron density of aromatic rings (benzene) of tyrosine & tryptophan in solution (pH 8)

A

ABSORPTION OF UV LIGHT AT 280 NM

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23
Q
  • anionic dye where albumin binds itself
A

BromcresolGreen Method (BCG)

24
Q

Albumin at pH [?] (acidic) is [?] (positive charge) & binds w/ [?]

A

4.2
cationic
anionic dye

25
Q

Dye-binding Methods used in spectrophotometric determination of serum albumin

A

(Bromcresol Green or Bromcresol purple)

26
Q

DYE-BINDING METHODS

Other dyes:

A

Bromcresol purple
Methyl orange
HABA(2 4-dihydroxyazobenzenebenzoic acid)
PSP (Phenolsulfonphthalein

27
Q

Detection of protein fractions after electrophoresis

A

DYE-BINDING METHODS

28
Q

TURBIDIMETRY & NEPHELOMETRY

 Precipitating agents

A

Sulfosalicylic acid (SSA) with sodium sulfate
Trichloroacetic acid

29
Q
  • to facilitate precipitation of both albumin and globulins
A

Sulfosalicylic acid (SSA) with sodium sulfate

30
Q

are often used for measuring urine & CSF protein

A

Turbidimetric methods

31
Q

[?] pH: protein donates protons = [?] charge

A

Alkaline
(-)

32
Q

Migrates towards ANODE

A

AlkalinepH: protein donates protons = (-) charge

33
Q
A

Acidic pH
(+) charge

34
Q

Migrates towards CATHODE

A

Acidic pH: protein it receives protons = (+) charge

35
Q

Typicalfor serum electrophoresis

A

Migrates towards ANODE

36
Q

No fibrinogen (consumed during coagulation)

A

Migrates towards ANODE

37
Q

No fibrinogen

A

AlkalinepH: protein donates protons = (-) charge

38
Q

 migrates the fastest toward the anode due to its low molecular
weight

A

Albumin

39
Q

Most anodal

A

Albumin

40
Q

Highest peak (most abundant protein in plasma)

A

Albumin

41
Q

The [?] manifest the concentration of the protein present (directly proportional)

A

peak of protein fraction

42
Q

Ex. Gamma globulin

A

Acidic pH: protein it receives protons = (+) charge

43
Q

Ex. Albumin

A

Alkaline pH: protein donates protons = (-) charge

44
Q

Protein stains after electrophoresis

A

Coomassie Brilliant Blue
Nigrosin dye
Ponceau S dye

45
Q

causes gamma globulins to migrate toward the cathode even though they are slightly (-) charged

A

ELECTROENDOSMOSIS

46
Q

due to electrical charge on support medium

A

ELECTROENDOSMOSIS

47
Q

Influences the migration of gamma globulins towards the cathode

A

electrical charge on support medium

48
Q

Relative concentration of each band determined by

A

densitometry

49
Q

plasma samples result in fibrinogen peak between [?]

A

gamma and beta fractions

50
Q

: due to IgA; Cirrhosis

A

Beta-gamma bridging effect (in serum)

51
Q

: due to Fibrinogen

A

Beta-gamma bridging effect (in plasma)

52
Q

: Monoclonal gammopathy (Multiple myeloma)

A

Monoclonal band (gamma globulin)

53
Q

: Chronic inflammation

A

Polyclonal band

54
Q

: nephrotic syndrome

A

Increase in a-2-macroglobulin

55
Q

: deficiency in a-1-antitrypsin; Juvenile cirrhosis

A

a-1-globulin flat curve

CC1 LECTURE DEAN (14 decks)

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