Rabbit White (GFAP) ELISA Test Company is a leading provider of high-quality enzyme-linked immunosorbent assay (ELISA) kits and related reagents. Our products are designed for efficient, sensitive, specific, and reliable detection of various analytes in biological samples such as body fluids, tissue homogenates, cell culture supernatants, and urine. All our reagents are offered at competitive prices, with exclusive discounts available to our customers. We also provide free test kits for scientific research purposes, specifically for the quantitative detection of human anti-neutrophil cytoplasmic antibody (cANCA) in cell lysates, body fluids, tissues, serum, plasma, and other sample types.
**Preparation of Materials and Reagents:**
1. **Instruments and Supplies:**
- Microplate reader (preheated for 30 minutes before use)
- Microtiter plates
- Pipettes and tips
- Distilled or deionized water
- Filter paper
2. **Reagent Preparation:**
- Dilute the working solution at a ratio of 1:30 according to the kit instructions.
**Sample Collection, Processing, and Storage:**
1. **Cell Culture Supernatant:**
Collect the supernatant using sterile tubes, centrifuge at 1000 × g for 15 minutes, and carefully transfer the supernatant for testing.
2. **Cells:**
Wash cells three times with PBS, adjust the concentration to approximately 10â´â€“10ⶠcells/mL. Lyse the cells by repeated freezing and thawing or ultrasonic disruption, then centrifuge to collect the supernatant.
3. **Serum:**
Allow blood to clot at room temperature, then centrifuge at 1000 × g for 15 minutes to obtain the serum.
4. **Plasma:**
Use EDTA or sodium citrate as an anticoagulant. Mix gently for 10–20 minutes, then centrifuge at 1000 × g for 15 minutes to collect the plasma.
5. **Body Fluids:**
Include pleural fluid, ascites, cerebrospinal fluid, and other secretions. Centrifuge the samples at 1000 × g for 15 minutes and collect the supernatant.
6. **Tissue Homogenate:**
Prepare by adding 9 mL of PBS (0.1 M, pH 7.4) to 1 g of tissue, thoroughly homogenize, and centrifuge to collect the supernatant. Avoid using NaN3, as it may inhibit horseradish peroxidase (HRP).
**Storage Recommendations:**
If immediate testing is not possible, aliquot the samples and store them at -70°C to prevent repeated freeze-thaw cycles. If precipitation occurs during storage, re-centrifuge the sample before use. Avoid hemolyzed or lipemic samples, and if the serum contains particulates, filter or centrifuge before testing. Do not thaw samples at 37°C or higher; instead, allow them to thaw at room temperature to ensure even and complete thawing.
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