Overview of Human Anticardiolipin Antibody IGM (ACA-IGM) ELISA Kit[1][2]
The human anticardiolipin antibody IGM (ACA-IGM) ELISA kit uses a double-antigen sandwich enzyme-linked immunoassay (ELISA) to determine the human anticardiolipin antibody IgM (ACA-IgM) in the specimen. The purified antigen is coated on the microplate to make a solid-phase antigen, which can bind to the anticardiolipin antibody IgM (ACA-IgM) in the sample. After washing to remove unbound antibodies and other components, it is then combined with the HRP-labeled antigen. Combine to form an antigen-antibody-enzyme-labeled antigen complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. Use a microplate reader to measure the absorbance (OD value) at a wavelength of 450 nm and compare it with the CUTOFF value to determine the presence or absence of human anticardiolipin antibody IgM (ACA-IgM) in the specimen.
Human Anticardiolipin Antibody IGM (ACA-IGM) ELISA Kit Sample Processing and Requirements[1][2]
1. Serum: Blood coagulates naturally at room temperature for 10-20 minutes, and centrifuges for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If precipitation occurs during storage, centrifuge again.
2. Plasma: EDTA or sodium citrate should be selected as the anticoagulant according to the requirements of the specimen. After mixing for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If any precipitate forms during storage, it should be centrifuged again.
3. Urine: Collect in a sterile tube and centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If any precipitate forms during storage, centrifuge again. Thoracic, ascites, and cerebrospinal fluid are performed as a reference.
4. Cell culture supernatant: When detecting secreted components, collect it in a sterile tube. Centrifuge for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant. When detecting intracellular components, dilute the cell suspension with rubber plasticizer PBS (PH7.2-7.4) until the cell concentration reaches about 1 million/ml. Freezing and thawing with anti-stearic acid can destroy cells and release intracellular components. Centrifuge for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant. If precipitate forms during storage, centrifuge again.
5. Tissue specimens: After cutting the specimens, weigh them. Add a certain amount of PBS, pH7.4. Quickly freeze and store in liquid nitrogen for later use. The specimen still maintains a temperature of 2-8°C after thawing. Add a certain amount of PBS (PH7.4) and homogenize the specimen fully by hand or with a homogenizer. Centrifuge for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant. After packaging, one portion will be tested and the rest will be frozen for later use.
6. The specimens should be extracted as soon as possible after collection. The extraction should be carried out according to relevant literature. The experiment should be carried out as soon as possible after extraction. If the test cannot be carried out immediately, the specimen can be stored at -20°C, but repeated freezing and thawing should be avoided.
7. Samples containing NaN3 cannot be detected because NaN3 inhibits the activity of horseradish peroxidase (HRP).
