For quantitative determination of immunglobulin IgG in human serum or plasma.
PRINCIPLE OF THE METHOD
Anti human IgG antibodies when mixed with samples containing IgG form insoluble complexes. These complexes cause an absorbance change, dependent upon the IgG concentration of the patient sample,that can be quantified by comparison from a calibrator of known IgG concentration. REAGENTS R1 (DILUENT): Tris buffer 20 mmol/L,PEG 8000 , Ph 8.3. Sodium azide 0.95 g/L. R2 (ANTIBODY): Goat serum, anti human IgG .Ph 7.5. Sodium azide 0.95 g/l CALIBRATION (Optional) Serum Proteins Calibrator is recommended.
REAGENT PREPARATION
Reagents are ready to use.
REAGENT DETERIORATION
Presence of particles and turbidity.
ADDITIONAL EQUIPMENT
1. Spectrophotometer or photometer thermostable at 37ºC with a 600 nm (580-620) nm filter.
2. Serum Proteins Calibrator and Serum Proteins Control are recommended for manual and automated assay procedures.
PROCEDURE
Wavelength : 600 nm (580-620)
Working temperature : 37°C
Optical path : 1 cm
Assay type : Turbidimetry
Direction : Increasing
CALCULATION
Calculate the absorbance difference (A2-A1) of each point of the calibration curve and plot the values obtained against the IgG concentration of each calibrator dilution. IgG concentration in the sample is calculated by interpolation of its(A2-A1) in the calibration curve.
QUALITY CONTROL
Serum Proteins Control are recommended to monitor the performance of manual and automated assay procedures. Each laboratory should establish its own Quality control scheme and corrective actions if controls do not meet the acceptable tolerances.