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Mustard ELISA Kit-total is intended for the quantitative determination of mustard (Sinapsis alba, Brassica juncea, Brassica nigra) in raw as well as heat treated foodstuff.

Mustard ELISA Kit-total contains all the required reagents for extraction and quantitative determination of mustard proteins.

Time required for the sample preparation and extraction: 1 hour (10 samples)
Time required for ELISA determination: 2 hours 10 min
Limit of detection (LOD): 0.05 mg/kg (ppm)
Limit of quantification (LOQ): 0.5 mg/kg (ppm)
Calibration scale range: 0.5 – 15 mg/kg (ppm)
Expiration time: 18 months (at 2 – 8 °C)

The mustard plant belongs to the Cruciferae (Brassicaceae) family. Mustard used in food is often a mixture of seeds from two or more species of Brassicaceae, for example Sinapis alba L. (yellow mustard), Brassica nigra (black mustard), and Brassica juncea L. (oriental mustard). Mustard is consumed with a variety of foods, and often is present in various spices and sauces as an ingredient. Allergic reactions to mustard, including severe anaphylactic reactions, are well documented in clinical and laboratory studies. In terms of its prevalence, mustard allergy is some fifth or sixth most frequent food allergy. Its incidence is closely related to mustard consumption in the region. The main allergens in mustard are proteins with the molecular weight of 14 kDa and 16 kDa marked as Sin a 1 and Bra j 1. These allergens are thermal-resistant and not subject to enzymatic cleavage. Minimal doses of Sin a 1 and Bra j 1 proteins to induce an allergic reaction are in microgram amounts.

The determination of mustard is based on its immunochemical reaction with a specific antibody. Mustard present in calibrator, control sample or analysed sample – extract of the analysed foodstuff - reacts in the first step with a specific antibody coated on walls of wells of a microtiter plate. Washing out of unbound proteins follows next the second, incubation step, during which the specific antibody conjugated with the enzyme – horse-radish peroxidase – reacts with the coated mustard. After expiry of the necessary incubation period comes washing out the wells and then the addition of a chromogenic substrate will enable to detect the remaining coated peroxidase. The intensity of colouration thus developed is proportional to the concentration of mustard in calibrators and samples.