Serum Albumin Determination

Jef Marc Valencia · 73問 · 2年前

通報

問題一覧

l. Mix well by _and stand at _ for_

GENTLY INVERSION, ROOM TEMPERATURE, 1 MIN

Read absorbance at_against the reagent blank.

600 NM

STABILITY

60 MIN

Determination of Albumin in serum or plasma is usually based on the binding behavior of the protein with an anionic dye

BCG METHOD

Excessive hemolysis:

INCREASED ALBUMIN

Color reagent has a reduced sensitivity to albumin-in-the-presence of

DETERGENT, DIOXANE

interferes with BCG Methods forming acidic pH

AMPICILLIN

At aneutral pH and low albumin concentration BCG wilbind with some of the _ and _ globulin present in human serum

ALPHA, BETA

When a solution containing proteins is rejected with a tartrate-complex cupric ions and_alkali_the_copper binds to the_peptide bond structure of the proteins.

MODIFIED BIURET TEST

Total Protein + Copper =

VIOLET COMPLEX

Modified Biuret Test Result

VIOLET COMPLEX

Linearity:

10 g/dL

Determination of amino Nitrogen derived from protein it uses serum treated with tungstic acid, forming PFF

KJELDAHL METHOD

KJELDAHL METHOD Nitrogen is converted to , using ,

NH3, H2SO4

TOTAL PROTEIN Specimen of choice: Interferences: Reference Value: -for ambulatory adults -in the recumbent position

SERUM, LIPEMIA, HEMOLYSIS, 6.5-8.3 g/dL, 6.0-7.8 g/dL

1gof Nitrogen = of PROTEIN

6.54 g

KJELDAHL METHOD Reference Method:

15.1%-16.8%

The nitrogen in ammonta may be measured using.

NESSLER REACTION, BERTHELOT REACTION, TITRATION METHOD

double iodide of Hg and K)

NESSLER REACTION

Nessler's Reagent

GUM GHATTI, AMMONIUM DIMERCURIC CHLORIDE

In Berthelot Reaction NH3 + AIkaline hypochlorite forms

INDOPHENOL BLUE

Copper binds to the peptide bond it requires at least 2 peptide bonds and an alkaline medium to measure TP

BIURET METHOD

BIURET METHOD positive result

PURPLE CHROMAGEN

oxidation of phenolic compounds such as tyrosine and tryptophan reduce PT-PMA reagent

FOLIN CIOCACTEU, LOWRY METHOD

(folin-ciocalteu) reagent

PHOSPHOTUNGSTIC MOLYBDIC ACID

LOWRY METHOD color enhancer

BIURET REAGENT

LOWRY METHOD Detects protein as low as

10-60 ug/mL

Widely used in research to measure tissue & enzyme proteins ' Not satisfactory for urine & CSF protein determination

LOWRY METHOD

The absorbance of proteins at 210 nm is due to the absorbance of the peptide bands at specific wavelength

ABSORPTION OF UV LIGHT AT 280 NM

28O NM absorption is due to

TRYPTOPHAN, TYROSINE, PHENYLALANINE

Proteins to bind certain dyes "Used to stain protein bands after electrophoresis (e.g. Coomassie Brilliant Blue)

DYE BINDING METHOD

Albumin at pH 4.2 is cationic & binds w/ anionic dye

BCG METHOD

Most specific dye for electrophoresis

BROMCRESOL PURPLE

Most commonly used dye

METHYL ORANGE

Precipitating agents: *SSA with sodium sulfate "Trichloroacetic acid

TURBIDIMETRY, NEPHELOMETRY

methods often used for measuring urine and CSF protein

TURBIDIMETRY

separates proteins on the basis of their electric charge densities

PROTEIN ELECTROPHORESIS

protein donates protons =(-) charge "Migrates fowards ANODE (+)

ALKALINE pH

protein receives protons =(+) charge Migrates towards CATHODE (-)

ACIDIC pH

confirms Multiple Myeloma:

GAMMA SPIKE

ALPHA I

ALPHA 1 ANTITRYPSIN, HDL, TGB, ALPHA 1 LIPOPROTEIN

ALPHA 2

HAPTOGLOBIN, CERULOPLASMIN, ALPHA 2 MACROGLOBULIN

BETA

TRANSFERRIN, BETA LIPOPROTEIN

Relative concentration of each band is determined by

DENSITOMETRY

BETA- GAMMA Bridging effect: due to IgA (in serum);

CIRRHOSIS

Monoclonal band (gamma-globulin) signifies

MONOCLONAL GAMMOPATHY

Polyclonal Band signifies

CHRONIC INFLAMMATION

Increase in a-2- macroglobulin:

NEPHROTIC SYNDROME

a-1-globulin flat curve

JUVENILE CIRRHOSIS, A-1-ANTITRYPSIN DEFICIENCY

plasma level above renal threshold as a result of metabolic disorder

OVERFLOW

enzyme deficiency which leads to: increased phenylalanine in the blood increased phenyl compounds in the urine

PHENYLKETONURIA

(branched chain ketoaciduria) oincreased branched chain amino acids in the blood and urine

MAPLE SYRUP URINE DISEASE

normal plasma level but decreased renal threshold or reabsorption

RENAL

increased _cystine, lysine ornithine, arginine in urine

CYSTINURIA

a, TLC with ninhydrin b. urine color tests c. Guthrie microbiological test. PKU

SCREENING TEST

a, Ion-exchange chromatography b. HPLC c. GC-MS

QUANTITATIVE TEST

COMPH Lesson 5

COMPH Lesson 5

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