Penicillin, BioAssay(TM) ELISA Kit

Penicillin was accidentally detected by Alexander Fleming in 1929. The drug belongs to the mycotoxins and is generated by the mould Penicillium chrysogenum. Penicillin as an antibiotic is preferentially used in the treatment of gram-positive bacteria, both for humans and animals. Of all illegally administered drugs, antibiotics are most frequently used in productive livestock. Contaminations in food or milk are ingested by humans, and can lead to severe infections by pathogen germs which became resistant against penicillin, or to allergies. The allergic reactions appear with different severity, dependent on dose and individual disposition, and showing symptoms from urticaria to anaphylactic shock. During routine testing of milk samples for antibiotics, in more than 90% of the positive cases, betalactam preparations or penicillins are detected.\n\nThe method of choice for the determination of penicillin contamination in food has always been a microbiological assay. These procedures allow however no quantitative determination and no identification of the antibiotic drug, which is achieved by a sensitive ELISA test kit or immunoaffinity columns together with HPLC.\n\nIntended Use:\nFor the quantitative determination of penicillin in food.\n\nSensitivity: \n3ng/ml\n\nRecovery (milk): ~90%\n\nRecovery (shrimp): ~70%\n\nIncubation Time: 140 min\n\nTest Principle:\nThe United States Biological Penicillin quantitative test is based on the principle of the enzyme linked immunosorbent assay. A penicillin conjugate is bound on the surface of a microtiter plate. Penicillin containing samples or standards and an antibody directed against penicillin are given into the wells of the microtiter plate. Immobilized and free penicillin compete for the antibody binding sites. After one hour incubation at room temperature, the wells are washed with diluted washing solution to remove unbound material. A peroxidase conjugate directed against the penicillin antibody is given into the wells and after another hour incubation, the plate is washed again. Then a substrate solution is added and incubated for 20 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 penicillin is indirectly proportional to the color intensity of the test sample.\n\nKit Components:\nP3175-20A: Microtiter Strips, 1x96 wells each, Coated with penicillin conjugate.\nP3175-20B: Standard 0ng/ml ,1x1ml, ready to use.\nP3175-20C: Standard 4ng/ml, 1x1ml, ready to use.\nP3175-20D: Standard 10ng/ml, 1x1ml, ready to use.\nP3175-20E: Standard 40ng/ml, 1x1ml, ready to use.\nP3175-20F: Standard 100ng/ml, 1x1ml, ready to use.\nP3175-20G: Standard 400ng/ml, 1x1ml, ready to use.\nP3175-20H: Anti-Penicillin Antibody (mouse), 1x6ml, ready to use.\nP3175-20J: IgG (HRP), 1x15ml, ready to use.\nP3175-20K: Substrate Solution (TMB), 1x15ml, ready to use.\nP3175-20L: Stop Solution, 0.5M H2SO4, 1x15ml, ready to use.\nP3175-20M: Sample Diluent 10X, (PBS), 2x50ml. \nP3175-20N: 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 at 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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SPECIFICATIONS

Catalog Number

P3175-20

Size

1Kit

Applications

ELISA

References

1. De Leuw P, Kapa G, Petz M, Schreurs FJ, Kan CA; J AOAC Int. 80(6): 1220 (1997). Induction and characterization of multianalyte antibodies against penicillins in egg yolk. 2. Knecht BG, Strasser A, Dietrich R, Martlbauer E, Niessner R, Weller MG; Anal Chem. 76(3): 646 (2004). Automated microarray system for the simultaneous detection of antibiotics in milk. 3. Althaus RL, Molina MP, Rodriguez M, Fernandez N; J Food Prot. 64(11): 1844 (2001). Detection limits of beta-lactam antibiotics in ewe milk by penzym enzymatic test. 4. Huth SP, Warholic PS, Devou JM, Chaney LK, Clark GH; J AOAC Int. 85(2): 355 (2002). Parallux beta-lactam: a capillary-based fluorescent immunoassay for the determination of penicillin-G, ampicillin, amoxicillin, cloxacillin, cephapirin, and ceftiofur in bovine milk. 5. Martlbauer E, Usleber E, Schneider E, Dietrich R; Analyst. 119(12): 2543 (1994). Immunochemical detection of antibiotics and sulfonamides. 6. Usleber E, Lorber M, Straka M, Terplan G, Martlbauer E; Analyst. 119(12): 2765 (1994). Enzyme immunoassay for the detection of isoxazolyl penicillin antibiotics in milk. 7. Gustavsson E, Sternesjo A; J AOAC Int. 87(3): 614 (2004). Biosensor analysis of beta-lactams in milk: comparison with microbiological, immunological, and receptor-based screening methods.