The FIV ELISA Kit is designed based on a double-antibody one-step sandwich ELISA method. This technique involves pre-coated microwells with feline immunodeficiency virus (FIV) capture antibodies. After adding the samples, standards, and HRP-labeled detection antibodies in sequence, the plate is incubated, washed thoroughly, and then developed using TMB substrate. Under peroxidase activity, TMB turns blue and then yellow when acid is added. The intensity of the color correlates directly with the amount of FIV present in the sample. The optical density (OD) at 450 nm is measured using a microplate reader to determine the sample concentration.
Sample collection and preparation are crucial for accurate results. For serum, collect blood in pyrogen-free tubes, centrifuge at 3000 rpm for 10 minutes, and separate the serum from red blood cells. Plasma should be collected using anticoagulants like EDTA, citrate, or heparin, followed by centrifugation at 3000 rpm for 30 minutes. Cell supernatant requires centrifugation at 3000 rpm for 10 minutes to remove debris. Tissue homogenate is prepared by chopping the tissue in saline and centrifuging at 3000 rpm for 10 minutes. Store samples in single-use aliquots at -20°C to avoid repeated freezing and thawing. Thaw samples at room temperature before use.
For optimal performance, ensure all reagents are well mixed before use. The kit components include microwells, standard solutions, detection antibody-HRP, wash buffer, substrates, stop solution, and sealing films. Prepare the 20× wash buffer by diluting it with distilled water in a 1:20 ratio. Manual washing involves five cycles of filling, soaking, and drying. Automatic washing uses 350 μL per well. Follow the protocol step-by-step: equilibrate the plate, add standards, samples, and detection antibody, incubate, wash, develop, stop, and read the OD values.
The kit offers high accuracy (R ≥ 0.99), sensitivity (<1.0 ng/ml), and specificity. It performs consistently with a coefficient of variation <15% between plates. Store the kit at 2–8°C, away from light and moisture, and use within 6 months. The detection range spans from 15.6 ng/ml to 500 ng/ml.
This kit is for research purposes only. Not suitable for clinical use or in vivo experiments. Users must strictly follow the instructions. Do not mix different batch numbers. The manufacturer is not liable for any errors resulting from improper handling. Always validate results and interpret them carefully.
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