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Casein ELISA Kit

Full Name: Casein ELISA Kit
Sample Type: Food (Soy Products, Bread Crumbs, Sausage, Chocolate, Cookies, Wine, Orange Juice)
Sensitivity: 0.04 ppm

BACKGROUND

Casein belongs to a family of phosphoproteins (αS1, αS2, β, κ) that are making up to 80% of the protein found in cow’s milk and approx. 20-45% of proteins present in human milk. The word casein comes from the Latin word caseus (meaning cheese), this is due the fact it represents a major ingredient in the production of cheese. It is frequently used as a food additive, types of glue (adhesive for labelling plastic bottles), in artists paint and by the dentist an agent in the re-minerlaisation of tooth enamel. Our bodies need to break down the casein to extract these peptides and amino acids.

There are three types found in cow’s milk: αS1, αS2 and β. The age-old controversy is whether the β-casein protein of cow’s milk is increasing the risk of developing diabetes. The primary function of casein in milk is to provide a source of calcium and phosphorous for infants. It also provides some protein (4%) for infant growth. Caseins are responsible for forming curd (tiny clumps) that form when milk is heated. In contrast to whey proteins which denature into a liquid at high temperatures, casein proteins are insoluble at all temperatures.

INTENDED USE

Casein ELISA kit is a procedure intended for quantitative measurement of bovine casein in food (such as bread crumbs, chocolate, cookies, soy products, sausage, orange juice and wine). This assay has a minimum analytical sensitivity limit of 0.04 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 anti-casein antibodies.
  • Casein Standards 1-5 (100x): Concentration 0, 0.2, 0.6, 2, 6 ppm.
  • Conjugate (Anti-Casein-Peroxidase).
  • Substrate Solution (TMB).
  • Stop Solution (0.5 M H2SO4).
  • Extraction And Sample Dilution Buffer (Carbonate Buffer)(5x).
  • Washing Solution (10x Concentrate).
  • Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of bovine casein using current casein ELISA kit was 0.04 ppm. The standard range for this assay is 0.2 – 6.0 ppm.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (0.04ppm), Limit of quantification, LOQ (0.2ppm)
– Specificity: The following displayed some cross reactions: Ewe’s milk (< 1.2%), Goat’s milk (< 1.2%). No cross reaction was detected for the following: Almond, Hazelnut, Pumpkin seed, Barley, Lecitihin, Rice (brown), Beef, Lima bean, Rice (white), Brazil nut, Oats, Rye, Buckwheat, Pea, Sacharose, Chicken, Peanut, Sesame, Chickpea, Pecan nut, Shrimps, Cocoa, Pineseed, Soy, Coconut, Pistachio, Split peas, Cod, Poppy seed, ß-Lactoglobulin, Corn, Pork, Walnut, Egg, Prawn, Wheat.
– Intra-Assay Precision: 5 – 11%
– Inter-Assay Precision: 8 – 14%
– Linearity: 100 – 127%
– Recovery: White wine (102%), Cookies (100%), Soy milk (94%), Chocolate (86%), Orange juice (84%), Bread crumps (80%), Sausage (80%).

REFERENCES

  1. Development of a sensitive ELISA for the detection of casein-containing fining agents in red and white wines. J Agric Food Chem. (2014) 62 (28): 6803-12. Deckwart M., et al.
  2. A simple and fast detection method for bovine milk residues in foods: a 2-site monoclonal antibody immunochromatography assay. J Food Sci. (2013) 78 (3): M452-7. Xuli W., et al.
  3. Specific antibodies in oral immunotherapy for cow’s milk allergy: kinetics and prediction of clinical outcome. Int Arch AllergyImmunol. (2014) 164 (1): 32-9. Savilahti E.M., et al.
  4. Digestibility and allergenicity assessment of enzymatically crosslinked beta-casein. Mol Nutr Food Res. (2010) 54 (9): 1273-84. Stanic D., et al.
  5. Allergenicity of alpha-caseins from cow, sheep, and goat. Allergy. (1997) 52 (3): 293-8. Spuergin P., et al.
  6. 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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