Folate Elisa Test Kit

INTENDED USE

The Folate ELISA kit is an enzymeimmunoassay intended for the quantitative determination of Folate in human serum, plasma, tissue homogenates and other biological fluids.

SUMMARY

Folate, distinct forms of which are known as folic acid, folacin, and vitamin B9,(1) is one of the B vitamins.(2)The recommended daily intake of folate in the US is 400 micrograms from foods or dietary supplements.(3) Folate in the form of folic acid is used to treat anemia caused by folic acid deficiency.(2) Folic acid is also used as a supplement by women during pregnancy to prevent neural tube defects (NTD) in the baby.(2,4)Low levels in early pregnancy are believed to be the cause of more than half of babies born with neural tube defects.(5) More than 50 countries use fortification of certain foods with folic acid as a measure to decrease the rate of NTDs in the population.(6,7) Long term supplementation is also associated with small reductions in the risk of stroke and cardiovascular disease. (8) It may be taken by mouth or by injection.(2)

Test principle

Competitive binding protein assay.

Total duration of assay: 75 minutes. The essential reagents required for a competitive binding assay include specific binding protein, enzyme-antigen conjugate and native antigen. Upon mixing enzyme-antigen conjugate, biotinylated binding protein and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of binding sites. A simultaneous reaction between the biotin attached to the binding protein and the streptavidin immobilized on the microwell occurs. This effects the separation of the binding protein enzyme bound fraction after decantation or aspiration. The enzyme activity in the protein binding protein bound fraction is inversely proportional to the native antigen concentration. By utilizing several different serum references of known antigen con- centration, a dose response curve can be generated from which the antigen concentration of an unknown can be

Reagents Materials provided

• Coated Microplate, 96 wells microplate coated with streptavidin.

• Calibrators, 6 vials, 1.0 mL each, ready to use; Concentrations: 0(A), 1.0(B), 2.5(C), 5.0(D), 10(E) and 25(F) ng/mL.

• Folate Enzyme Reagent: 1 vial, 6.0 mL. Contains Folate (Analog)-horseradish peroxides (HRP) conjugate in a protein-stabilizing matrix with dye. • Folate Biotin Reagent: 1 vial, 6.0 mL. Contains biotinylated purified folate binding protein conjugate in buffer, dye and preservative.

• Releasing Agent: 1 vial, 12 mL. Contains a strong base (sodium hydroxide) and potassium cyanide.

• Stabilizing Agent: 1 vial, 6 mL. Contains tris (2-carboxyethyl) phosphine (TCEP) solution.

• Neutralizing Buffer: 1 vial, 10 mL. Contains buffer with dye that reduces the pH of sample extraction.

• TMB Substrate, 1 vial, 11 mL, ready to use, (tetramethylbenzidine) TMB.

• Stop Solution, 1 vial, 6.0 ml of 1 mol/L sulfuric acid.

• Wash Solution Concentrate, 1 vial, 30 mL (20X concentrated), PBS-Tween wash solution.

• IFU, 1 copy

• Plate Lid, 1 piece

• Sealed Bag, 1 piece

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