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Beta-Lactoglobulin ELISA Kit (Beta-LG)

Full Name: Beta-Lactoglobulin ELISA Kit (Beta-LG)
Sample Type: Food (Soy Products, Chocolate, Cookies, Sausage, Cereals, Orange Juice, Wine)
Sensitivity: 1.5 ppm

BACKGROUND

Beta-Lactoglobulin (beta-LG) is present in ruminant species or the milk of many other species. It is main whey protein found in cow and sheep’s milk (approx. 3g/l), it belongs to the lipocalins family that has the ability to bind a large number of hydrophobic molecules (indicating a potential transport function). Lipocalin proteins also contain molecular pockets that allow them to accommodate iron complexes (siderohores) and organic molecules in a pH-dependent manner. Beta-Lactoglobulin are found in the milk of many mammalian species. The lipocalins family is composed of more than 30 proteins, with differing properties and functions. Lipocalins are glycoproteins that contain tryptophan, histidine, tyrosine and cysteine residues and carry one or two lactosyl residue (a disaccharide containing one molecule each of glucose and galactose).

The molecular mass of bovine beta-lactoglobulin is approx. 48.3 kDa, it circulates in the blood and is excreted by the kidneys in a resorbable form. It is found in humans’ blood, breast milk, and lungs. It is a component of normal plasma proteins, but it may be present in greater amounts in some people than others. Cow’s milk has been promoted as an infant formula or mother’s milk substitute for people with certain medical conditions that require a high protein diet due to its high content of globulin and trace minerals (such as iron).

INTENDED USE

Beta-lactoglobulin ELISA kit can be used to measuring quantitative amounts of ß-lactoglobulin (beta lactoglobulin, beta-LG, beta-lactoglobulin) in food samples (for example chocolate, cookies, cereals, soy products, sausage, wine and orange juice). This assay has a minimum analytical sensitivity limit of 1.5 ppm.

CONTETNT

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-beta-lacto-globulin antibodies.
  • Beta-Lactoglobulin Standards 1-5: Concentration 0, 10, 40, 100, 400 ppb.
  • Conjugate (anti-beta-lactoglobulin-peroxidase).
  • Substrate Solution (TMB).
  • Stop Solution (0.5 M H2SO4).
  • Extraction and sample dilution buffer (Tris)(10x).
  • Washing Solution (10x Concentrate).
  • Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of ß-lactoglobulin (beta-LG, beta lactoglobulin) using current beta-lactoglobulin ELISA kit was 1.5 ppm. The standard range for this assay is 10.0 – 400.0 ppm.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (1.5ppm), Limit of quantification, LOQ (10ppm)
– Cross Reactivity: The following displayed some cross reactions: Ewe’s milk (< 0.2%), Casein (< 0.02%), Goat’s milk (< 0.002%). No cross reaction was detected for the following: Egg, Poppy seed, Walnut, Chick pea, Peach, Wheat, Sesame, Pecan nut, Pea, Apricot, Oats, Sunflower seed, Brazil nut, Bean, Cherry, Rye, Pumpkin seed, Coconut, Lecithin, Plum, Barley, Pine nut, Almond, Saccharose, Beef, Rice, Cashew nut, Pistachio, Cocoa, Pork, Corn, Peanut, Macadamia, Orange, Chicken, Buckwheat, Hazelnut, Chestnut, Wine, Bovine albumin, Soy.
– Intra-Assay Precision: 7%
– Inter-Assay Precision: 9 – 12%
– Linearity: 72 – 127%
– Recovery: Sausage (107%), Orange juice (98%), Cereals (94%), Cookies (88%), Chocolate (86%), White wine (82%), Soy milk (70%).

REFERENCES

  1. Comparison of immunomodulating properties of Beta-lactoglobulin and its hydrolysates. Iran J AllergyAsthma Immunol. (2014) 13 (1): 26-32. Duan C.C., et al.
  2. ELISA testing for soy antigens in dry dog foods used in dietary elimination trials. J Am Anim Hosp Assoc. (2014) 50 (6): 383-9. Willis-Mahn C., et al.
  3. Food allergy: a winding road to the present. Pediatr AllergyImmunol. (2014) 25 (1): 25-6. Sampson H.A.
  4. Yogurt in the Treatment of β-Lactoglobulin-Induced Gastrointestinal Cow’s Milk Allergy. J Investig Allergol Clin Immunol. (2016) 26 (5): 327-329. Poza-Guedes P., et al.
  5. T-cell epitope-containing hypoallergenic β-lactoglobulin for oral immunotherapy in milk allergy. Pediatr AllergyImmunol. (2016) 27 (8): 818-824. Ueno H.M., et al.

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