VMA(Vanillylmandelic Acid) ELISA Kit

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

Product Details

Properties

Assay type	Competitive-ELISA
Format	96T
Assay time	2.0h
Detection range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes VMA in samples. No significant cross-reactivity or interference between VMA and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of VMA concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with VMA. During the reaction, VMA in the sample or standard competes with a fixed amount of VMA on the solid phase supporter for sites on the Biotinylated Detection Ab specific to VMA. 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 VMA 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 VMA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level VMA were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	4.60	10.90	45.90	4.80	11.80	49.40
Standard deviation	0.30	0.50	1.50	0.20	0.50	2.30
CV (%)	6.52	4.59	3.27	4.17	4.24	4.66

Recovery

The recovery of VMA 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)	86-98	91
EDTA plasma(n=8)	93-105	99
Cell culture media(n=8)	85-99	90

Target Information

Research Area

Cancer, Metabolism

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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