Sample
Plasma (ACD- or heparinized) and Serum
Intended Use
A solid-phase enzyme immunoassay for the quantitative determination of CA72-4 in human serum or plasma. This kit is designed for measurement of CA72-4 in blood serum or plasma. The kit contains reagents sufficient for 96 determinations and allows to analyze 42 unknown samples in duplicates.
Contents of Kit
CA72-4 EIA strips: 96 (8x12) wells
Polystyrene microwells coated with murine monoclonal Ab to human CA72-4.
Calibrator set (1-5): 0.5 mL x 5, the set contains 5 calibrators: 0; 5; 15; 50; 200 U/mL.
Human CA72-4 diluted in a preselected human serum preservative - 0.01% Bronidox L, 0.01% 2-Methyl-4-isothiazolin-3-one-hydrochloride; also contains blue dye. Ready-to-use. (C1-colourless)
Control serum: Ready-to-use. 0.5 mL
Dilution of preselected human serum, with high content of CA72-4 with preservative - 0.01% Bronidox L, 0.01% 2-Methyl-4-isothiazolin-3-one-hydrochloride; also contains purple dye.
Conjugate concentrate (11x): 1.2 mL
Aqueous solution of murine monoclonal to human CA72-4 coupled with horseradish peroxidase diluted on phosphate buffered saline with casein from bovine milk and detergent (Tween-20), contains 0.1% phenol as preservative and green dye.
Conjugate dilution buffer: 12 mL
Aqueous Tris-buffered BSA solution, preservative-0.01% Bronidox L, 0.01% 2-Methyl-4-isothiazolin-3-one-hydrochloride; contains light blue dye.
Substrate solution: 14 mL
Ready-to-use single-component tetramethylbenzidine (TMB) solution.
Washing solution concentrate 26x: 22 mL
Aqueous solution of sodium chloride and detergent (Tween 20), contains proClin300 as a preservative.
Stop solution: 14 mL
5.0% vol/vol solution of sulphuric acid. Ready-to-use.
Plate sealing tape 2 slides
Storage
Store the whole kit at 2 to 8°C upon receipt until the expiration date. A short-term storage at higher temperatures (nmt 25°C for nmt 5 days) is allowed (e.g., during transportation). After opening the pouch keep unused microtiter wells TIGHTLY SEALED BY ADHESIVE TAPE (INCLUDED) to minimize exposure to moisture.
Stability of opened/diluted components
CA72-4 EIA strips: until exp. date
Calibrator set (Calibrator 1-5): 2 months
Control serum: 2 months
Conjugate concentrate: concentrate - until exp. date; diluted - 1 day at 2-8°C
Conjugate dilution buffer: until exp. date
Substrate solution: until exp. date
Washing solution concentrate 26x: concentrate - until exp. date; diluted washing solution - 45 days at 2-8°C or 15 days at RT
Stop solution: until exp. date
Plate sealing tape: N/A
Sensitivity
Sensitivity of the assay was assessed as being 1.0 U/mL.
General Description
CA 72-4, or a carbohydrate antigen 72-4, is a high MW (230-1000 kD) antigen (epitope) associated to gastric and ovarian cancer as well as some other malignancies and not expressed in noticeable quantities in tissues of healthy adult individuals. Quantitative determination of CA 72-4 in serum or plasma is helpful (particularly, in combination with CA 19-9) for monitoring of gastric cancer and its therapy, while combined determination of CA 72-4 and CA 125 is used for monitoring of ovarian cancer.
Elevated levels of CA 72-4 are often seen in adenocarcinomas of the gastro-intestinal tract, ovaries (mucinous type) and lungs. Besides, raised CA 72-4 is sometimes also seen in persons with benign pathology (chronic inflammation, cysts, fibrosis). That is why, results of CA 72-4 determination should always be interpreted in conjunction with other laboratory and clinical data.
NOTE: the individuals that have received murine monoclonal antibodies for radioimaging or immunotherapy may develop high titered anti-mouse antibodies (HAMA). The presence of these antibodies gives false results in the present assay. Sera from HAMA positive persons should be treated by depleting adsorbents before assaying.
Standard Curve
Below is presented a typical example of a standard curve. Not for calculations!
Citations
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