FA/VB9(Folic Acid/Vitamin B9) ELISA Kit

Detection Range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL

Product Details

Properties

Assay Type	Competitive-ELISA
size	96T
Assay Time	2.5h
Detection Method	Colormetric
Detection Range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL
Sample Volume	50μL
Sample Type	serum, plasma and other biological fluids
Specificity	This kit recognizes Universal FA/VB9 in samples.No significant cross-reactivity or interference between Universal FA/VB9 and analogues was observed
Precision	Both intra-CV and inter-CV are < 10%.
Recovery	80%-120%
Introduction	This ELISA kit applies to the in vitro quantitative determination of FA/VB9 concentrations in serum, plasma and other biological fluids.
Applications	ELISA
Storage	2-8℃/-20℃

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal FA/VB9. During the reaction, Universal FA/VB9 in the sample or standard competes with a fixed amount of Universal FA/VB9 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal FA/VB9. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal FA/VB9 in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips	-20℃, 6 months
Micro ELISA Plate(Dismountable)	48T: 8 wells ×6 strips
Reference Standard	96T: 2 vials
Reference Standard	48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL
Concentrated Biotinylated Detection Ab (100×)	48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL	-20℃(Protect from light), 6 months
Concentrated HRP Conjugate (100×)	48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and
high level FA/VB9 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and
high level FA/VB9 were tested on 3 different plates, 20 replicates in each plate,
respectively.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	5.4	8.46	35.72	4.86	8.06	38.99
Standard deviation	0.37	0.48	1.96	0.28	0.36	2.09
CV (%)	6.85	5.67	5.49	5.76	4.47	5.36

Recovery

The recovery of FA/VB9 spiked at three different levels in samples throughout the
range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	85-100	91
EDTA plasma(n=8)	90-101	95
Cell culture media(n=8)	101-117	108

Target Information

Research Area

Cell Biology, Metabolism, Immunology, Developmental Biology, Biochemicals

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

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ELISA Assays