Technical Data
Almond, BioAssay™ ELISA Kit (Prunus dulcis)
Kits and Assays Storage: 4°CShipping: Blue Ice
Almond (Prunus dulcis) belongs to the rosaceae. With 25 % the fraction of proteins in almond is high. Some of these proteins are known for being allergenic. In addition to profilin which is known to be cross-reactive to grass pollen, the almond major protein AMP has the greatest relevancy. AMP is known to be heat resistant making it stable to different production processes. For this reason almond represents an important food allergen. For almond allergic persons hidden almond allergens in food are a critical problem. Already very low amounts of almond can cause allergic reactions, which may lead to anaphylactic shock in severe cases. Because of this, almond allergic persons must strictly avoid the consumption of almonds or almond containing food. Cross-contamination, mostly in consequence of the production process is often noticed. The chocolate production process is a representative example. This explains why in many cases the existence of almond residues in foods cannot be excluded. For this reason sensitive detection systems for almond residues in foodstuffs are required. The United States Biological Almond ELISA represents a highly sensitive detection system and is particularly cap-able of the quantification of almond residues in cookies, cereals, ice cream and chocolate.

Intended Use:
For the quantitative determination of almond in food.


Recovery (spiked samples): ~85-95%

Incubation Time: 200 min

Test Principle:
The United States Biological Almond quantitative test is based on the principle of the enzyme linked immunosorbent assay. An antibody directed against almond prote-ins is bound on the surface of a microtiter plate. Almond containing samples or standards are given into the wells of the microtiter plate. After 20 minutes incubation at room temperature, the wells are washed with diluted washing solution to remove unbound material. A peroxidase conjugated second antibody directed against almond proteins is given into the wells and after 20 minutes of incubation the plate is washed again. A substrate solution is added and incubated for 20 minutes, resulting in the devel-opment of a blue color. The color development is inhibited by the addition of a stop solution, and the color turns yel- low. The yellow color is measured photometrically at 450nm. The concentration of almond is directly proportional to the color intensity of the test sample.

Kit Components:
1. Microtiter plate 1x96 wells each, coated with anti-almond antibodies.
2. Standard 0ng/ml 1x1ml, ready to use.
3. Standard 0.4ng/ml 1x1ml, ready to use.
4. Standard 1ng/ml 1x1ml, ready to use.
5. Standard 4ng/ml 1x1ml, ready to use.
6. Standard 10ng/ml 1x1ml, ready to use.
7. Conjugate (anti-almond-peroxidase): 1x15ml, ready to use.
8. Substrate Solution (TMB): 1x15ml, ready to use.
9. Stop Solution (0.5M H2SO4): 1x15ml, ready to use.
10. Extraction and sample dilution buffer 10x (Tris): 2x120ml.
7. Washing Solution 10x (PBS + Tween 20): 1x60ml.
Dilute 1+9 with distilled water. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.

Storage and Stability:
Store components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
1. Roux KH, et al. (2001) - Detection and stability of the major almond allergen in foods. J Agric Food Chem, 49(5):2131-6. 2. Venkatachalam M, et al. (2002) - Effects of roasting, blanching, autoclaving and microwave heating on antigenicity of almond (Prunus dulcis L.) proteins. J Agric Food Chem, 50(12):3544-8. 3. Ben Rejeb S, et al. (2005) - Multi-allergen screening immunoassay for the detection of protein markers of peanut and four tree nuts in chocolate. Food Addit Contam, 22(8):709-15. 4. Acosta MR, et al. (1999) - Production and characterization of rabbit polyclonal antibodies to almond (Prunus dulcis L.) major storage protein. J Agric Food Chem, 47(10):4053-9. 5. Tawde P, et al. (2006) - Cloning and charcterization of profilin (Pru du 4 ), a cross-reactive almond (Prunus dulcis) allergen. J Allergy Clin Immun, 118(4):915-922. 6. De Leon MP, et al. (2003) - Immunological analysis of allergenic cross-reactivity between peanut and tree nuts. Clin Exp Allergy, 33(9):1273-80. 7. Shridar K, et al. (2001) - Electrophoretic and immunological analyses of almond (Prunus dulcis L.) genotypes and hybrids. J Agric Food Chem, 49(4):2043-52. 8. Shridar K, et al. (2002) - Biochemical characterization of amandin, the major storage protein in almond (Prunus dulcis L.). J Agric Food Chem, 50(15):4333-41. 9. Gaur V, et al. (2008) - Purification, identification and preliminary crystallographic studies of pru du amandin, an allergenic protein from Prunus dulcis. Acta Cryst Sect Struct Biol Cryst Com, 64(1):32-5. 10. Lee SH, et al. (2005) -A 50 kDa maize gamma-zein has marked cross reactivity with the almond major protein. J Agric Food Chem, 53(20):7965-70.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.