FMN (Flavin mononucleotide) ELISA Kit

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SKU:
AEES00665

Description

Product Name:FMN (Flavin mononucleotide) ELISA Kit
Product Code:AEES00665
Assay Type:Competitive
Format:96T
Assay Time:2.5h
Detection Range:15.63-1000 pg/mL
Sensitivity:9.38 pg/mL
Sample Volume:50μL
Specificity:This kit recognizes FMN in samples. No significant cross-reactivity or interference between FMN 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 FMN concentrations in serum, plasma and other biological fluids.

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with FMN. During the reaction, FMN in the sample or standard competes with a fixed amount of FMN on the solid phase supporter for sites on the Biotinylated Detection Ab specific to FMN. 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 FMN in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level FMN were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level FMN were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)54.07150.26353.1249.95141.68339.22
Standard deviation2.978.6112.643.256.8912.45
CV (%)5.495.733.586.514.863.67

Recovery

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

Sample TypeRange (%)Average Recovery (%)
Serum(n=8)89-10697
EDTA plasma(n=8)87-10092
Cell culture media(n=8)93-107100
Kit Components:

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.

ItemSpecificationsStorage
Micro ELISA Plate(Dismountable)8 wells x 12 strips-20℃, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100×)1 vial, 120 μL
Concentrated HRP Conjugate (100×)1 vial, 120 μL-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL2-8°C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25×)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL2-8℃(Protect from light)
Stop Solution1 vial, 10 mL2-8℃
Plate Sealer5 pieces
Manual1 copy
Certificate of Analysis1 copy
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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Additional Information

Product Type:
ELISA Kit
ELISA Type:
Competitive
Size:
96 Assays
Sensitivity:
9.38 pg/mL
Range:
15.63-1000 pg/mL
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