ELISA

Microbial Transglutaminase (MTG) ELISA


ELISA for the detection of Microbial Transglutaminase (MTG)

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1 Kit 425 € available


Documents
Art. No. E021
Background info Microbial transglutaminase catalyzes acyl transfer reactions between the γ-carboxyamide group of a protein-bound glutamine residue and primary amines. In simple terms, microbial transglutaminase acts as a biological glue that covalently links proteins. Among others microbial transglutaminase is used in the food industry for the production of dairy products, meat, sausages and bakery products.

The Microbial Transglutaminase (MTG) ELISA (Product Nr. E021) is a sandwich enzyme immunoassay developed for the quantitative determination of native MTG. As part of the kit development, the following foods were examined: glued meat (raw), yoghurt, quark, cheese, salami and bread. It can be assumed that the test is also suitable for the analysis of further foods. Detection and determination limits depend on the respective sample matrix, the degree of processing and the extraction process.

Please note that for each type of sample material, the preparation procedure must be validated separately by the user.

All reagents for the implementation of the enzyme immunoassay are contained in the test kit. A test kit is sufficient for 96 determinations (including standards). A microtiter plate photometer is required for evaluation.

Sample preparation: homogenizing, extracting
Standard material: Microbial Transglutaminase (0 - 150 ng/mL)
Detection limit: 0.6 ng/mL (ppb) MTG (1 mL ≙ 1 g)
Determination limit: 3.0 ng/mL (ppb) MTG (1 mL ≙ 1 g)

MTG ELISA
Assay principle The microtiter plate wells (solid phase) are coated with antibodies specific to microbial transglutaminase.
1st reaction: Microbial transglutaminase present in the sample binds to the immobilised antibodies, forming the antibody-antigen-complex.
2nd reaction: A microbial transglutaminase specific antibody labeled with biotin binds to the antibody-antigen-complex forming an antibody-antigen-[antibody-biotin] sandwich.
3rd reaction: Streptavidin conjugated to horseradish peroxidase (HRP) binds to the biotin-label fo the antibody-antigen-[antibody-biotin] sandwich.
4th reaction: The HRP-labeled complex converts a substrate (TMB) into a blue product which upon addition of the stop solution turns yellow. Samples containing microbial transglutaminase develop the blue colour (which upon addition of the stop solution turns yellow), whereas samples without microbial transglutaminase remain colourless.
Reagents in the kit Microtiter plate packed in a resealable foil laminate pouch with a dessicant bag
Sample buffer
Wash buffer
MTG-standard.
Purified anti-microbial transglutaminase antibody labeled with biotin (MTG-Ab conjugate)
Streptavidin-HRP conjugate
Substrate solution
Stop solution (0.2 M H2SO4)
Directions for use
Storage Store the whole kit at 6 - 8°C. It is important that the MTG-standard is not stored below 6°C. The kit is stable up to the expiry date stated on the label of the box. Do not use kit beyond its expiry date. After opening the pouch keep unused microtiter wells resealed to minimize exposure to moisture. Substrate solution is sensitive to light, therefore avoid direct exposure to light. After the expiry date (see kit label) no quality guarantee is given.
Note INTENDED FOR RESEARCH USE ONLY, NOT FOR USE IN HUMAN, THERAPEUTIC OR DIAGNOSTIC APPLICATIONS.

Artikel in DLG-Lebensmittel – 06/2017 zur Bestimmung von Transglutaminase in Lebensmitteln (deutsch)

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