Folic Acid ELISA

$680.00

SKU: FOL06-K01 Categories: , ,
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Folic Acid ELISA Assay kit:
For Research Use Only
Size:  1×96 wells
Sensitivity:  2 ng/mL
Dynamic Range:  4 – 400 ng/mL
Incubation Time:  2 hours
Sample Type:  Food Products, Cell Culture, Tissue
Sample Size: 3 – 10 g

Product Developed and Manufactured in Germany
Product Support in the USA

Intended Use
The Eagle Biosciences Folic Acid ELISA Assay kit is intended for the quantitative determination of Folic Acid (Vitamin B9) in food products by enzyme linked immunoassay (ELISA).  The Eagle Biosciences Folic Acid ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

Assay Principle
The Folic Acid ELISA Assay kit is a quantitative assay based on the principle of the enzyme linked immu­nosorbent assay. A folic acid conjugate is bound on the surface of a microtiter plate. Folic acid containing samples or standards and an antibody directed against folic acid are given into the wells of the microtiter plate. Immobi­lized and free folic acid compete for the antibody binding sites. After one hour incubation at room tem­perature, the wells are washed with diluted washing solution to remove unbound material. A peroxidase conjugate against the antibody is given into the wells and after another hour incubation, the plate is washed again. Then a substrate solution is added and incubated for 20 minutes, resulting in the devel­opment of a blue colour. The colour development is inhibited by the addition of a stop solution, and the colour turns yellow. The yellow colour is measured photometrically at 450 nm. The concentration of folic acid is indirectly proportional to the colour inten­sity of the test sample.

  1. Pipet 100 µL standards or prepared samples in duplicate into the appropriate wells of the micro­titer plate. Immediately add 50 µL folic acid antibody into each well.
  2. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
  3. Wash the plate three times as follows: Dis­card the contents of the wells (dump or aspi­rate). Pipet 300 µL of diluted washing solu­tion into each well. After the third repetition empty the wells again and remove residual liquid by striking the plate against a paper towel. The wash procedure is critical. In­suf­fi­cient washing will result in poor precision and falsely ele­vated absorbencies.
  4. Pipet 100 µL of conjugate (anti-mouse-IgG-HRP) into each well.
  5. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
  6. Wash the plate as outlined in #3.
  7. Pipet 100 µL of substrate solution into each well.
  8. Allow the reaction to develop in the dark (e.g. cupboard or drawer; the chromogen is light-sensi­tive) for 20 minutes at room temperature.
  9. Stop enzyme reaction by adding 100 µL of stop solution (0.5 M H2SO4) into each well. The blue colour will turn yellow upon addition.
  10. After thorough mixing, measure absorbance at 450 nm (reference wavelength 620 nm), using an ELISA reader. The colour is stable for 30 minutes.


Assay Background
Folates play an important role in the synthesis of nucleic acids and some amino acids and gained re­cently increased interest because they belong to the group of antioxidative vitamins. In the last years the in­fluence of folic acid supplementation to avoid abortion and dysraphism was a topic of research increasingly. Folic acid as the most stable representative of the group of folates is added to a broad range of food.

Traditional methods are mostly microbiological ones, but also TLC and HPLC are applied. These methods are time consuming and need complex equipment.  The Eagle Biosciences Folic Acid ELISA test kit allows the detection (2.5 to 4 hrs. incl. sample preparation) of folic acid in supplemented food which is more rapid compared to traditional techniques (24-48 hrs).

References:

  • Pfeiffer, C. et al, Z. Ernährungswiss., 33, 85-119 (1994).
  • Rothenberg, S.P. et al, New England J. of Medicine, 286, 1335-1339 (1972).
  • Dunn, R.T., Foster, L.B., Clin. Chem., 19/10, 1101-1105 (1973).
  • Shane, B., Clin. Chim. Acta, 100, 13-19 (1980).
  • Müller, H., Z. Lebensm. Unters. Forsch., 196, 518-521 (1993).

Recent Citations:

Kantar, MB;Anderson, JE;Lucht, SA;Mercer, K;Bernau, V;Case, KA;Le, NC;Frederiksen, MK;DeKeyser, HC;Wong, ZZ;Hastings, JC;Baumler, DJ;, (2016). Vitamin Variation in Capsicum Spp. Provides Opportunities to Improve Nutritional Value of Human Diets. PLoS ONE, 11(8), e0161464