Milk ELISA Kit

Full Name: Milk ELISA Kit
Sample Type: Food (Cookies, Chocolate, Bread Crumbs, Soy, Sausage, Wine, Orange Juice)
Sensitivity: 0.05 ppm

BACKGROUND

Milk is usually processed into dairy products for example butter, cheese, yogurt, cream and ice-cream. It can also be found in many other found such as bread, breakfast cereals, processed meats, ready meals, soups, pizza, puddings and custards, sweets and cakes to juts name a few. A milk allergy, is very much like most of the other food allergies, it can occur during early infancy and cows’ milk protein allergy is regarded as the most frequent, however, many children tend to grow out of this allergy after the age of 5. Although most children do grow out of cow’s milk protein allergy, it can occur in people of any age. In these cases, there is a missed or delayed diagnosis and the cause is unknown. The most common causes of allergy in children are cow’s milk and soy-based formula products which contains soy protein.

The following assay offers a very sensitive method for analysing milk based on NIST 1549 reference material, this protocol is validated to quantify casein and ß-lactoglobulin residues present in foods such as sausage, soy products, bread crumbs, cookies, chocolate, wine and orange juice. An individual that it allergic to milk should therefore avoid milk or even the presence of hidden milk proteins present in foods such as convenience food, cookies, sausage and beverages. Approx. 80 % of bovine milk proteins are caseins and also beta-Lactoglobulin, represents a major allergen of whey (this represents an additional 10% of the total protein). In order to measure bovine milk in foodstuffs, a highly sensitive detection procedure is essential.

INTENDED USE

Milk ELISA kit is designed for measuring quantitative amounts of bovine milk protein present in food (for example chocolate, bread crumbs, cookies, soy products, sausage, orange juice and wine). This assay has a minimum analytical sensitivity limit of 0.05 ppm.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

Microtiter Plate: Coated with milk protein binding antibodies.
Milk protein Standards 1-5 (100x): Concentration 0, 0.4, 1, 4, 10 ppm. Based on NIST RM 1549 reference material.
Conjugate (Anti-Milk Protein-Peroxidase).
Substrate Solution (TMB).
Stop Solution.
Extraction and sample dilution buffer (Carbonate buffer)(5x concentrate).
Washing Solution (PBS + Tween 20), (10x Concentrate).
Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of bovine milk using current milk ELISA kit was 0.05 ppm. The standard range for this assay is 0.4 – 10.0 ppm.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (0.05ppm), Limit of quantification, LOQ (0.4ppm)
– Specificity (Cross Reactivity): Ewe’s milk (0.94%) Goat’s milk (0.01%). No cross reaction was detected for the following: Almond, Egg, Pork, Barley, Hazelnut, Rice, Beef, Lupin, Rye, Celery, Mustard, Sesame, Chicken, Oats, Soy, Cocoa, Peanut, Walnut, Cod, Pistachio, Wheat Corn.
– Intra-Assay Precision: 8 – 10%
– Inter-Assay Precision: 10 – 17%
– Linearity: 80 – 130%
– Recovery: White wine (122%), Bread crumbs (110%), Orange juice (106%), Cookies (102%), Chocolate (99%), Sausage (88%), Soy milk (79%).

REFERENCES

A rapid immunomagnetic beads-based immunoassay for the detection of β-casein in bovine milk. Food Chem. (2014) 158: 445-8. Song F., et al.
Short communication: Milk ELISA status for bovine leukosis virus infection is not associated with milk production in dairy cows. J Dairy Sci. (2011) 94 (10): 5062-4. Sorge U.S., et al.
Role of maternal elimination diets and human milk IgA in the development of cow’s milk allergy in the infants. Clin Exp Allergy. (2014) 44 (1): 69-78. Järvinen K.M., et al.
Human milk_proteins may interfere in ELISA measurements of bovine beta-lactoglobulin in human milk. Acta Paediatr. (1996) 85 (5): 543-9. Bertino E., et al.
Commercial Milk Enzyme-Linked Immunosorbent Assay (ELISA) Kit Reactivities to Purified Milk-Proteins and Milk-Derived Ingredients. J Food Sci. (2016) 81 (7): T1871-8. Ivens K.O., et al.

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