Product Specification

        Carbohydrate antigen 153 (CA153) Tumor Markers

INTENDED USE

   Detection Kit for Carbohydrate antigen 15-3(CA153)(ELISA). It is used in quantitative tests for CA15-3 in human serum.

SUMMARY AND CLINICAL SIGNIFICANCE

    Carbohydrate antigen 15-3 (CA15-3) is a epitope fragment MUC-1of breast cancer cell, which is a glycoprotein. CA15-3 present in a variety of cancer, such as the breast, lung cancer and ovarian cancer. CA15-3 is used for determining breast cancer progression and metastasis, and clinical testing treatment and relapse in Clinical. It has been reported that when CA15-3> 100kU / L, the metastatic breast cancer lesions can be considered. When the treatment is effective about 50% of patients CA15-3 concentrations were significantly decreased; Concentration of CA15-3 are more sensitive than radiological and clinical criteria in 96% local recurrence. This reagent mainly is used for dynamic monitoring of cancer patients to determine disease progression or auxiliary therapeutic effect, but its concentration is not correlated with tumor size, growth, and the degree of malignancy grading / staging. In some benign diseases (Such as benign breast disease, cirrhosis, hepatitis, etc.) can also be seen that the index increased in varying degrees. Therefore, the CA15-3 should not be considered an effective general screening test for the general population of cancer screening, early diagnosis and other purposes.

PRINCIPLE

CA15-3 uses a “sandwich principle”, Enzyme-linked immunological sorbent assay. To measure CA15-3 levels in serum, plastic wells coated with a monoclonal antibodies of CA15-3 are supplied in the kit. The patient’s specimen is added and, during incubation, CA15-3, if present, is fixed to the solid phase antibody, when mono-antibody labelled with HRPis added, creating a HRP-antibody—CA15-3—antibody “sandwich”. After added TMB substrate added, the result is obtained by EIA plate reader.

REAGENTSSUPPLIED

Coated Microplate: 1 plate (8 × 12 wells), Ready to use. Coated with anti-CA15-3 antibody and sealed in an aluminum bag. Remove the strips in the resealable bag with a desiccant to protect from moisture after opened. Store at 2~8℃ until expiration date. 

HRP Conjugate: 1 vial of 11ml, Ready to use. Containing HRP labeled anti-CA15-3 and proclin-300 preservative. Store at 2~8℃ until expiration date.

Sample dilution: 1 vial of 11ml, Ready to use. Store at 2~8℃ until expiration date.

Calibrator:5 vials of 0.5ml, Ready to use. Labeled with S0 to S4 and the concentration of CA-15-3 is 0,15,40,100,250 U/ml. Store at 2~8℃ until expiration date.

Control: 1 vial of 0.5ml, Ready to use. Store at 2~8℃ until expiration date. The concentration is 53~98 U/ml.

Chromogen A: 1 vial of 7ml, Ready to use. Containing H2O2. Store at 2~8℃ until expiration date.

Chromogen B: 1 vial of 7ml, Ready to use. Containing TMB. Store at 2~8℃ until expiration date.

Stop Solution: 1 vial of 7ml, Ready to use. Containing H2SO4. Store at 2~8℃ until expiration date.

Wash buffer: 1 vial of 15ml, Concentrate 20-fold, diluting with deionized water before the assay. Containing NaCl, phosphate and tween-20. Store at 2~8℃ until expiration date.

Plate sealer: 2 pieces.

Plastic resealable bag: 1 set.

Instruction manual: 1 copy.

PREPARATIONS

1.  Allow all specimens and reagents to reach room temperature and mix thoroughly by gentle inversion before use.

2. Prepare Wash Buffer by diluting Wash Concentrate 20-fold with deionized water. The diluted wash solution is stable in room temperature for at least one week.

ASSAY PROCEDURE:

1.Mark the microtitration strips to be used. All the calibrator and controls should set duplicate.

2.Dispense 100 ul of calibrator/controls. Dispense 100 ul of sample dilution and 10 ul sample into respective wells.

3.Covered the strips with a plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 0.5hour.

4.Wash each well for 3 times, 10 seconds each time. (See wash procedure)

5. Dispense 100 ul of HRPConjugateto each well.

6. Covered the strips with a plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 0.5 hour.

7.Wash each well for 3 times, 10 seconds each time. (See wash procedure).

8.Dispense 50 ul of chromogen Ato each well.

9.Dispense 50 ul of chromogen B to each well.

10.Covered the strips with a fresh plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 15 minutes.

11.Dispense 50 ul ofstoppingsolutionto each well andmix completely.

12. Readtheabsorbanceof theplate within 10 minutes. (See read procedure)

PRECAUTION FOR USERS

1.   Handling should preclude any pipetting by mouth. Use only pipettes with disposable tips for each specimen.

2.Do not mix materials from different master lots. Do not use kit components beyond the expiration date.

3.All materials should be brought to room temperature before use.