CDSimple™ Folate ELISA Kit

Name: CDSimple™ Folate ELISA Kit
SKU: DEIACL4
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Serum

Species Reactivity

Human

Detection Method

cELISA

Intended Use

The Quantitative Determination of Folate Concentration in Human Serum by a Microplate Enzyme Immunoassay, Colorimetric

Contents of Kit

A. Folate Calibrators - 1.0 ml/vial
Six (6) vials containing human serum albumin reference for Folate at concentrationsof 0(A),1.0(B),2.5(C),5.0(D),10.0 (E), and 25.0 (F) in ng/ml. A preservative has been added. Store at 2-8°C
Note: The calibrators, human serum based, were calibrated using a highly purified N_methyltetrahydrofolate preparation.
B. Folate Enzyme Reagent -7.0 ml/vial
One (1) vial containing Folate (Analog)-horseradish peroxides (HRP) conjugate in a protein-stabilizing matrix with dye. Store at 2-8°C
C. Folate Biotin Reagent-7.0 ml/vial
One (1) vial containing biotinylated purified folate binding protein conjugate in buffer, dye and preservative. Store at 2-8°C
D. Streptavidin Coated Plate- 96 wells
One 96-well microplate coated with streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8°C
E. Wash Solution Concentrate -20.0 ml/vial
One (1) vial containing a surfactant in buffered saline. A preservative has been added. Store at 2-8°C.
F. Substrate Reagent - 12.0 ml/vial
One (1) vial containing tetramethylbenzidine (TMB) and hydrogen peroxide(H₂O₂)in buffer. Store at 2-8°C.
G. Stop Solution -8.0 ml/vial
One (1) vial containing a strong acid (0.5M H₂SO₄). Store at 2-8°C.
H. Releasing Agent -14.0 ml/vial
One (1) vial containing a strong base (sodium hydroxide) and potassium cyanide. Store 2-8°C.
I. Stabilizing Agent-0.7 ml/ vial
One (1) vial containing tris(2-carboxyethyl) phosphine (TCEP) solution.Store at 2-8°C
J.Neutralizing Buffer-7.0 ml/ vial
One (1) vial containing buffer with dye that reduces the pH of sample extraction. Store at 2-8°C
K. Product Insert
Note 1: Do not use reagents beyond the kit expiration date.
Note 2: Avoid extended exposure to heat and light. Opened reagents are stable for sixty (60) days when stored at 2-8°C. Kit and component stability are identified on the label.
Note 3: Above reagents are for a single 96-well microplate.

Storage

2-8°C

Performance Characteristics

Accuracy
The Folate ELISA Test System was compared with a reference method. Biological specimens were used with values that ranged from 3.2ng/ml - 13.7ng/ml. The total number of such specimens was 30. The least square regression equation and the correlation coefficient were computed for this Folate ELISA in comparison with the reference method. The data obtained is displayed in Table.

Only slight amounts of bias between this method and the reference method are indicated by the closeness of the mean values. The least square regression equation and correlation coefficient indicates excellent method agreement.

Precision

The within and between assay precision of the Folate ELISA Test System were determined by analyses on three different levels of pool control sera. The number, mean values, standard deviation and coefficient of variation for each of these control sera are presented in Tables.

*As measured in ten experiments in duplicate over a ten day period.

Sensitivity

The Folate ELISA Test System has a sensitivity of 0.52ng/ml. The sensitivity was ascertained by determining the variability of the 0 ng/ml serum calibrator and using the 2б (95% certainty) statistic to calculate the minimum dose.

General Description

Folate supplementation has escalated over recent years with the knowledge of its many benefits. As one of the B vitamins, folate, or Vitamin B9, is involved in many bodily functions and deficiency can cause disease in not only the elderly but infants too. Folate deficiency is associated with megaloblastic anemia, neural tube defects, and cardiovascular dieases.
Folate plays an important role in brain development and therefore is vital during growth. The most common defects resultant from folate deficiencies are neural tube defects. With a vital role in nucleic acid synthesis, folate has been found to be beneficial as supplementation during pregnancy and other times of rapid tissue growth. Folate also plays a vital role in maintaining proper balance of homocysteine, a contributing factor in occurrences of occlusive vascular diseases and stroke. Individuals with susceptibility to heart disease and several forms of cancer may also benefit from supplementation.
Major sources of folate include green leafy vegetables, legumes, beans and fortified cereals. Foods fortified with folate are actually fortified with folic acid because of the higher bioavailability for absorption by the body. In circulation, folate is present in several different forms, some of which are more stable than others. Folic aicd and methyltetrahydrofolate are two common forms, the latter being more stable and found in higher concentrations in serum.
Due to the stability of the molecule, methytetrahydrofolate is very often used as the form focused on during methods of
analysis.
Folate binding proteins are responsible for folate metabolism. Two types exist in circulation; one type aids in binding to the cell surface and the other soluble form exists in circulation. These folate binding proteins also have the capability of binding several different folate derivatives including folic acid and methytetrahydrofolate. The interaction between folic acid and folate binding protein is greater than methyltetrahydrofolate. Current assays on the market require an extraction step to release the folate derivatives from the folate binding protein.
In the past, folate has been quantified in samples using such methods as microbiological assays, bio-specific procedures and HPLC-MS techniques. Overall, this rapid rise in knowledge of folate, its importance, and subsequently folate supplementation has caused a higher demand for improved testing methods.

Standard Curve