Histamine, BioAssay(TM) ELISA Kit

Histamine is a biogenic amine, which is formed by enzymatical decarboxylation from the amino acid histidine. It occurs in the mast cells and basophilic white cells bound to heparin. In the course of a type I allergy, endogenic histamine from the mast cells and basophilic leukocytes is released after antigen binding to membrane-associated IgE, and typical allergical reactions appear.\n\nBut histamine can also enter the human body via the nutrition and can thus cause pseudoallergic food intolerances. The appearing symptoms are not distinguishable from those of a real allergy. The term 'food allergy' is however only used, if the pathologic symptoms develop in the course of an immunological reaction. Non-immunological reactions to food are defined as food intolerances. Food intolerances, which are caused by increased histamine concentrations, are clinically characterized by rash, diarrhoea, vomiting, nausea, itching, headache and asthma. The extent of the reaction is dependent on the ingested amount of histamine. These reactions can be enhanced by a lack of diaminoxidase.\n\nSeparated from histamine intolerances, toxic reactions exist, which are caused by very high histamine concentrations. Toxic histamine concentrations, which can cause the so-called scombroid reaction, appear after bacterial degradation of protein-rich food, especially fish. Histamine is found above all in cheese, fish, spinach, smoked products and red wine. The histamine concentrations in different types of food, but also in the same food show frequently great differences. In different types of cheese there can e.g. be found histamine contents between 600 and 1000mg/kg. \n\nHistamine ELISA is a quick, economical and sensitive method to detect histamine in wine, fish or cheese. After an appropriate sample preparation, 40 samples can be tested in duplicate within 80 minutes.\n\nIntended Use:\nFor the quantitative determination of Histamine in food.\n\nSensitivity: 2ng/mL\n\nRecovery (wine, fish, cheese): ~90%\n\nIncubation Time: 80 min\n\nTest Principle:\nHistamine, Food, BioAssay(TM) ELISA Kit quantitative test is based on the principle of the enzyme linked immunosorbent assay. A histamine conjugate is bound on the surface of a microtiter plate. Samples or standards containing derivatized histamine and an antibody directed against histamine are given into the wells of the microtiter plate. Immobilized and free histamine compete for the antibody binding sites. After 30 minutes incubation at room temperature, the wells are washed with diluted washing solution to remove unbound material. A peroxidase conjugate directed against the histamine antibody is given into the wells and after another 30 minutes 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 histamine is indirectly proportional to the color intensity of the test sample.\n\nKit Components:\n1. Microtiter plate 1x96 wells each, coated with histamine conjugate.\n2. Standard 4ng/ml 1x4ml each.\n3. Standard 2ng/ml 1x4ml each.\n4. Standard 4ng/ml 1x4ml each.\n5. Standard 10ng/ml 1x4ml each.\n6. Standard 40ng/ml 1x4ml each.\n7. Standard 100ng/ml 1x4ml each.\n8. Reaction Solution: 1x3ml, ready to use.\n9. Neutralizing Solution: 1x15ml, ready to use.\n10. Anti-Histamine Antibody (mouse): 1x6ml, ready to use.\n11. Conjugate (anti-mouse-IgG-HRP): 1x15ml, ready to use.\n12. Substrate Solution (TMB): 1x15ml, ready to use.\n13. Stop Solution (0.5 M H2SO4): 1x15ml, ready to use.\n14. Sample Diluent (PBS): 1x60ml, ready to use.\n15. 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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