ACA IgM ELISA
For in vitro diagnostic and professional use only.
INTENDED USE
This kit is a qualitative detection of human serum/plasma of Anticardiolipin IgM antibody. The kit is suitable for clinical screening and diagnosis.
ACA antibody is a group of auto-antibodies against a variety of negatively charged phospholipids, It can be used as one of the diagnostic criteria for antiphospholipid antibody syndrome (including thrombosis), spontaneous abortion, thrombocytopenia and central nervous system (CNS) lesions. ACA IgM can be used as prospective index of spontaneous abortion; Positive ACA was significantly associated with CNS thrombosis in patients with systemic lupus erythematosus (SLE).
| Product details |
Description |
| Delivery |
Within 48 hours |
| Packaging Specifications |
8 x 12 strips, 96 wells |
| Country Of Origin |
China |
| Manufacturer |
18 months |
| Preservation method |
2℃-8℃ |
| Specimen |
Whole blood |
| Assification |
class1 |
| Type |
Elisa Test Kit |
PRINCIPLE
This kit uses indirect ELISA principle to detect ACA IgM. Purified ACA antigen is pre-coated on the microplate, the ACA IgM antibody in sample will combine with ACA antigen first, then combine with enzyme-labeled second antibody to form antigen-antibody-antibody complex, and shows blue color in the microplate. This kit is used for the specific detection of ACA IgM in human serum/plasma.
TEST PROCEDURE
1. All reagents should be allowed to reach room temperature for 15 minutes before use.
2. Dilute the wash buffer at the rate of 1:40 dilution with distilled water before use.
3. Add 100μL Sample Diluent in the corresponding well, add 5μL sample in the corresponding well (Do not add in the blank well). Mix thoroughly by using the pipette. Add 50μL of positive control and negative control to the positive control well and negative control well. The sample should be corresponding to the number of micro plate, each plate should be provided with negative control 2 wells, positive control 1 well and blank control 1 well. Note: Use a separate disposal pipette tip for each specimen, Negative and Positive Control to avoid cross-contamination.
4. Shake gently to mix for 30s. Incubate at 37°C for 20 minutes with the sealing plate membrane sealing the plate.
5. At the end of the incubation, remove and discard the plate cover. Take out, add wash buffer to each well for 20 seconds. Repeat 5 times. After the final washing cycle, turn the plate over onto blotting paper or clean towel, and tap it to remove any remainders.
6. Respectively adding Conjugate 50μL (Do not add in the blank well) .
7. Incubate at 37°C for 20 minutes with the sealing plate membrane sealing the plate. Repeat the wash step for 5 times as in step 5.
8. Add Substrate A (50µL) and Substrate B (50µL) (Do not add in the blank well). Incubate at 37°C for 10 minutes with the sealing plate membrane sealing the plate.
9. Add 50μL Stop Solution to each well (Do not add in the blank well). Mix gently by shaking, read the absorbance within 10 minutes after stopping the reaction. Calibrate the plate reader with the Blank well and read the absorbance at 450nm. If a dual filter instrument is used, set the reference wavelength at 630nm. Set no blank wells is allowed if use dual wavelength to detect. Calculate the Cut-off value and evaluate the results.
Storage
• Store at 2-8℃.
• Place unused wells in the zip-lock aluminum foiled pouch and return to 2-8 °C, under which conditions the wells will remain stable for 2 months, or until the labeled expiry date whichever is earlier.
• Seal and return unused calibrators to 2-8 °C, under which conditions the stability will be retained for 1 month, for longer use, store opened calibrators in aliquots and freeze at-20 °C. Avoid multiple freeze-thaw cycles.
• Seal and return all the other unused reagents to 2-8 °C, under which conditions the stability will be retained for 2 months, or until the labeled expiry date, whichever is earlier.
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