Bovine PAEP ELISA Kit

Sample

Food

Detection Method

sELISA

Intended Use

Enzyme Immunoassay for the Quantitative Determination of β-Lactoglobulin in Food.

Contents of Kit

The kit contains reagents for 96 determinations. They have to be stored at 2-8°C. Expiry data are found on the labels of the bottles and the outer package.
1. Microtiter plate consisting of 12 strips with 8 breakable wells each, coated with anti-β-lactoglobulin antibodies.
2. β-Lactoglobulin Standards (0, 10, 40, 100, 400 ppb of βlactoglobulin): 5 vials with 1.0 mL each, dyed red, ready-touse.
3. Conjugate (anti-β-lactoglobulin-peroxidase): 15 mL, dyed red, ready-to-use.
4. Substrate Solution (TMB): 15 mL, ready-to-use.
5. Stop Solution (0.5 M H₂SO₄): 15 mL, ready-touse.
6. Extraction and sample dilution buffer (Tris): 2 × 120 mL as 10× concentrate, dyed red. Dilute 1+9 with distilled water. Stored at 4°C the diluted buffer is stable for at least one week. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.
7. Washing Solution (PBS + Tween 20): 60 mL as 10× concentrate. Dilute 1+9 with distilled water. Stored at 4°C the diluted buffer is stable for at least 4 weeks. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.
8. Plastic bag to store unused microtiter strips.
9. Instruction Manual.

Storage

Stored at 2-8°C. Expiry data are found on the labels of the bottles and the outer package. For more detailed information, please download the following document on our website.

Precision

Intra-assay Precision: 7%
Inter-assay Precision: 9 - 12%

Sensitivity

The limit of detection (LOD) of the β-Lactoglobulin test is 1.5 ppb. Validation experiments with milk (products) showed that this corresponds approximately to 0.6 ppm unskimmed milk, 0.1 ppm non fat milk powder (NIST RM1549) and 0.5 ppm whole milk powder (NIST RM8435). The limit of quantification (LOQ) of the β-Lactoglobulin test is 10 ppb. Validation experiments with milk (products) showed that this corresponds approximately to 3.7 ppm unskimmed milk, 0.6 ppm non fat milk powder (NIST RM1549) and 3.2 ppm whole milk powder (NIST RM8435). Due to the variety of sample matrices and their influence on the blank, results less than the LOQ should be treated as negative.

General Description

Bovine milk belongs to the most important allergenic food ingredients especially for children. Already very low amounts of bovine milk can cause allergic reactions, which may lead to anaphylactic shock in severe cases. Because of this, milk allergic persons must strictly avoid the consumption of milk or milk containing food. In particular the presence of hidden milk proteins such as in sausage, cookies, convenience food or beverages represent a critical problem for milk allergic persons. According to EU Directive 2003/89/EG the addition of bovine milk has to be labeled. For the detection of bovine milk in foodstuffs sensitive detection systems are required. Approximately 20% of bovine milk proteins are whey proteins. The main fraction of whey proteins consists of the heat-stable allergen β-lactoglobulin. The β-Lactoglobulin ELISA represents a highly sensitive detection system and is particularly capable of the identification and quantification of bovine milk residues in cookies, cereals, sausage, orange juice, wine, soy products and chocolate.