Chloramphenicol, BioAssay(TM) ELISA Kit (Chloromycetin)

Due to its outstanding antibacterial properties, chloramphenicol is an often used antibiotic in the production of milk, meat and eggs. In humans it leads to haematotoxic side effects, like the chloramphenicol induced aplastic anemia. This has caused low limits in Germany, e.g. 1ug/kg for milk, meat and eggs.\n\nTill now the chloramphenicol concentration was determined by radioimmunoassay or by gas chromatography. However, compared with conventional methods, enzyme immunoassays show some essential advantages. There is no need to work with radioactive material, the required assay time is shorter and the sensitivity is better than with chromatographic methods.\n\nThe United States Biological Chloramphenicol test provides a rapid, sensitive and reliable assay for the determination of chloramphenicol in food. 40 samples can be assayed in duplicate within 60 minutes.\n\nIntended Use:\nFor the quantitative determination of chloramphenicol in food.\n\nSensitivity: \n0.03ng/ml\n\nRecovery (spiked samples): ~90-115%\n\nIncubation Time: 60 min\n\nTest Principle:\nThe United States Biological Chloramphenicol quantitative test is based on the principle of the enzyme-linked immunosorbent assay. An antibody binding protein is coated on the surface of a micro- titer plate. Chloramphenicol containing samples or standards and an antibody directed against chloramphenicol are given into the wells of the microtiter plate. The chloramphenicol contained in samples or standards will bind to the antibody which reacts with the binding protein coated onto the microtiter plate. After 30 minutes incubation at room temperature a chloramphenicol-peroxidase conjugate is added into the wells without a preceding washing step to saturate free antibody binding sites. After additional 15 minutes incubation at room temperature the wells are washed with diluted washing solution to remove unbound material. A substrate solution is added and incubated for 15 minutes, resulting in the development of a blue color. The color development is inhibited by the addition of a stop solution, and the color turns yellow. The yellow color is measured photometrically at 450nm. The concentration of chloramphenicol is indirectly proportional to the color intensity of the test sample.\n\nKit Components:\n1. Microtiter plate 1x96 wells each, coated with an antibody binding protein.\n2. Standard 0ng/ml 1x1ml, ready to use.\n3. Standard 0.05ng/ml 1x1ml, ready to use.\n4. Standard 0.1ng/ml 1x1ml, ready to use.\n5. Standard 0.5ng/ml 1x1ml, ready to use.\n6. Standard 1ng/ml 1x1ml, ready to use.\n7. Standard 5ng/ml 1x1ml, ready to use.\n8. Anti-Chloramphenicol Antibody (sheep): 1x6ml, ready to use.\n9. Conjugate (Chloramphenicol-Peroxidase): 1x6ml, ready to use.\n10. Substrate Solution (TMB): 1x15ml, ready to use.\n11. Stop Solution (0.5 M H2SO4): 1x15m ready-to-use.\n12. Sample Diluent (PBS): 2x60ml, ready to use.\n13. Washing Solution 10x (PBS + Tween 20): 1x60ml. \nDilute 1+9 with distilled water. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37 degrees C for 15 minutes.\n\nStorage and Stability:\nStore components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

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