Mustard, BioAssay(TM) ELISA Kit

Mustard belongs to the Brassica plant family. With about 30-35% the fraction of proteins in mustard seed is very high. Some of these proteins are known for being allergenic, such as Sin a 1 and Bra j 1. These proteins are predominantly heat resistant making them stable to different production processes. In addition to brown mustard (Brassica juncea) and black mustard (Brassica nigra) primarily yellow mustard (Sinapsis alba) is used as an ingredient in many foods and food preparations. For mustard allergic persons hidden mustard allergens in food are a critical problem. Already very low amounts of mustard can cause allergic reactions, which may lead to anaphylactic shock in severe cases. Because of this, mustard allergic persons must strictly avoid the consumption of mustard or mustard containing food. Cross-contamination, mostly in consequence of the production process, is often noticed. The sau-sage production process is a repre-sentative ex-ample. This explains why in many cases the exis-tence of mustard residues in food cannot be ex-cluded. For this reason sensitive detection systems for mustard residues in foodstuffs are required. The United States Biological Mustard ELISA represents a highly sensitive detection system for yellow mustard and is particularly capable of the quantification of residues in sausage, dressings, soups, cheese and mixed herbs. Due to high cross-reactivity the test is also suitable for the detection of brown mustard and black mustard.\n\nIntended Use:\nFor the quantitative determination of mustard in food.\n\nSensitivity: \n1ppm\n\nRecovery (spiked samples): ~76-98%\n\nIncubation Time: 60 min\n\nTest Principle:\nThe United States Biological Mustard quantitative test is based on the principle of the enzyme linked immunosorbent assay. An antibody directed against mustard proteins is bound on the surface of a microtiter plate. Mustard con- taining samples or standards are given into the wells of the microtiter plate. After 20 minutes incubation at room temperature, the wells are washed with diluted washing solution to remove unbound material. A peroxidase conjugated second antibody directed against mustard proteins is given into the wells and after 20 minutes of incubation the plate is washed again. 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 mustard is directly proportional to the color intensity of the test sample.\n\nKit Components:\n1. Microtiter plate 1x96 wells each, coated with anti-mustard antibodies.\n2. Standard 0ng/ml 1x1ml, ready to use.\n3. Standard 2ng/ml 1x1ml, ready to use.\n4. Standard 6ng/ml 1x1ml, ready to use.\n5. Standard 20ng/ml 1x1ml, ready to use.\n6. Standard 60ng/ml 1x1ml, ready to use.\n7. Conjugate (anti-mustard-peroxidase): 1x15ml, ready to use.\n8. Substrate Solution (TMB): 1x15ml, ready to use.\n9. Stop Solution (0.5M H2SO4): 1x15ml, ready to use.\n10. Extraction and sample dilution buffer 10x (Tris): 2x120ml. \n11. 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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SPECIFICATIONS

Catalog Number

M9545-01

Size

1Kit

Applications

ELISA

References

1. Lee PW, et al. (2008) - Sandwich enzyme-linked immunosorbent assay (ELISA) for detection of mustard in food. J Food Sci, 73(4):62-68. 2. Koppelmann SJ, et al. (2007) - Development of an enzyme-linked immunosorbent assay method to detect mustard protein in mustard seed oil. J Food Prot, 70(1):179-83. 3. Shim YY, et al. (2008) - Quantitative detection of allergenic protein Sin a 1 from yellow mustard (sinapsis alba L.) seeds using enzyme-linked immunosorbent assay. J Agric Food Chem, 56(4):1184-92. 4. Lee PW, et al. (2009) - Detection of mustard, egg, milk and gluten in salad dressing using enzyme-linked immunosorbent assays (ELISAs). J Food Sci, 74(5):46-50. 5. Palomares, et al. (2005)