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CC1 LECTURE DEAN > PROTEIN DETERMINATION > Flashcards

PROTEIN DETERMINATION Flashcards

(55 cards)

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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
Study These Flashcards
A

BromcresolGreen Method (BCG)

24
Q

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

Study These Flashcards
A

4.2
cationic
anionic dye

25
Dye-binding Methods used in spectrophotometric determination of serum albumin
(Bromcresol Green or Bromcresol purple)
26
DYE-BINDING METHODS Other dyes:
Bromcresol purple Methyl orange HABA(2 4-dihydroxyazobenzenebenzoic acid) PSP (Phenolsulfonphthalein
27
Detection of protein fractions after electrophoresis
DYE-BINDING METHODS
28
TURBIDIMETRY & NEPHELOMETRY  Precipitating agents
Sulfosalicylic acid (SSA) with sodium sulfate Trichloroacetic acid
29
- to facilitate precipitation of both albumin and globulins
Sulfosalicylic acid (SSA) with sodium sulfate
30
are often used for measuring urine & CSF protein
Turbidimetric methods
31
[?] pH: protein donates protons = [?] charge
Alkaline (-)
32
Migrates towards ANODE
AlkalinepH: protein donates protons = (-) charge
33
[?]: protein it receives protons =
Acidic pH (+) charge
34
Migrates towards CATHODE
Acidic pH: protein it receives protons = (+) charge
35
Typicalfor serum electrophoresis
Migrates towards ANODE
36
No fibrinogen (consumed during coagulation)
Migrates towards ANODE
37
No fibrinogen
AlkalinepH: protein donates protons = (-) charge
38
 migrates the fastest toward the anode due to its low molecular weight
Albumin
39
Most anodal
Albumin
40
Highest peak (most abundant protein in plasma)
Albumin
41
The [?] manifest the concentration of the protein present (directly proportional)
peak of protein fraction
42
Ex. Gamma globulin
Acidic pH: protein it receives protons = (+) charge
43
Ex. Albumin
Alkaline pH: protein donates protons = (-) charge
44
Protein stains after electrophoresis
Coomassie Brilliant Blue Nigrosin dye Ponceau S dye
45
causes gamma globulins to migrate toward the cathode even though they are slightly (-) charged
ELECTROENDOSMOSIS
46
due to electrical charge on support medium
ELECTROENDOSMOSIS
47
Influences the migration of gamma globulins towards the cathode
electrical charge on support medium
48
Relative concentration of each band determined by
densitometry
49
plasma samples result in fibrinogen peak between [?]
gamma and beta fractions
50
: due to IgA; Cirrhosis
Beta-gamma bridging effect (in serum)
51
: due to Fibrinogen
Beta-gamma bridging effect (in plasma)
52
: Monoclonal gammopathy (Multiple myeloma)
Monoclonal band (gamma globulin)
53
: Chronic inflammation
Polyclonal band
54
: nephrotic syndrome
Increase in a-2-macroglobulin
55
: deficiency in a-1-antitrypsin; Juvenile cirrhosis
a-1-globulin flat curve

CC1 LECTURE DEAN (14 decks)

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