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

Cat.No.:E-EL-0009
Reactivity: Universal

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96T
48T
24T
96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
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 FA/VB9 in samples. No significant cross-reactivity or interference between FA/VB9 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 FA/VB9 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 FA/VB9. During the reaction, FA/VB9 in the sample or standard competes with a fixed amount of FA/VB9 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 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 FA/VB9 in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & 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 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 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.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	5.40	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

Citations

Journal of Crohns & Colitis (2015) IF: 9.071
Diet-Induced Hyperhomocysteinaemia Increases Intestinal Inflammation in an Animal Model of Colitis

DOI: 10.1093/ecco-jcc/jjv094

Sample: Plasma
Nanoscale (2016) IF: 7.76
A TiS2 nanosheet enhanced fluorescence polarization biosensor for ultra-sensitive detection of biomolecules

DOI: 10.1039/C6NR00946H

Sample: serum
Oxidative Medicine and Cellular Longevity (2022) IF: 6.543
Mitochondrial DNA and Epigenetics: Investigating Interactions with the One-Carbon Metabolism in Obesity

DOI: 10.1155/2022/9171684

PMID: 35132354
Sample: Serum
Frontiers in Molecular Biosciences (2021) IF: 5.246
Folic Acid Alleviates High Glucose and Fat-Induced Pyroptosis via Inhibition of the Hippo Signal Pathway on H9C2 Cells

DOI: 10.3389/fmolb.2021.698698

PMID: 34692767
Sample: H9C2 cell
Scientific Reports (2017) IF: 4.259
Folate Metabolism Regulates Oligodendrocyte Survival and Differentiation by Modulating AMPKα Activity

DOI: 10.1038/s41598-017-01732-1

Sample: Serum
Nutrition Journal (2020) IF: 3.359
Low folate intake and serum levels are associated with higher body mass index and abdominal fat accumulation: a case control study.

DOI: 10.1186/s12937-020-00572-6

Sample: Serum
ANNALS OF NUTRITION AND METABOLISM (2021) IF: 3.374
Comparison of Associations between One-Carbon Metabolism, Lipid Metabolism, and Fatty Liver Markers in Normal-Weight and Overweight People Aged 20–40 Years

DOI: 10.1159/000517911

Sample: serum
NORDIC JOURNAL OF PSYCHIATRY (2019) IF: 2.061
Klotho, vitamin D and homocysteine levels during acute episode and remission periods in schizophrenia patients

DOI: 10.1080/08039488.2019.1582697

PMID: 30896269
Sample: Serum
Neuropsychiatric Disease and Treatment (2019) IF: 2.228
Comparison of serum B12, folate and homocysteine concentrations in children with autism spectrum disorder or attention deficit hyperactivity disorder and healthy controls

DOI: 10.2147/NDT.S212361

PMID: 31496704
Sample: Serum
AQUACULTURE NUTRITION (2015) IF: 1.395
Effects of dietary folic acid on growth, antioxidant capacity, non‐specific immune response and disease resistance of juvenile Chinese mitten crab Eriocheir sinensis (Milne‐Edwards, 1853)

DOI: 10.1111/anu.12275

Sample: Tissue homogenate
AQUACULTURE (2014) IF: 1.828
Dietary vitamin B12 requirement and its effect on non-specific immunity and disease resistance in juvenile Chinese mitten crab Eriocheir sinensis

DOI: 10.1016/j.aquaculture.2014.08.010

Sample: Tissue homogenate
Indian Journal of Clinical Biochemistry (2022)
Metabolic Analysis of Methylenetetrahydrofolate Reductase Single Nucleotide Polymorphisms (MTHFR 677C

DOI: 10.1007/s12291-022-01049-5

Sample: serum
African Health Sciences (2020) IF: 0.6899
Relationship between plasma homocysteine and blood pressure in hypertensive Northern-Nigerians

DOI: 10.4314/ahs.v20i1.38

Sample: plasma

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