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ELK Biotechnology

ELK9671

Mouse CNCbl(Cyanocobalamin) ELISA Kit

Mouse CNCbl(Cyanocobalamin) ELISA Kit

Sensitivity

Mouse CNCbl(Cyanocobalamin) ELISA Kit

Overview

Product name: Mouse CNCbl(Cyanocobalamin) ELISA Kit
Reactivity: Mouse
Alternative Names: VB12; Vitamin B12; Cobalamin; α-5,6-Dimethylbenzimidazolyl Cobamidcyanide
Assay Type: Sandwich
Sensitivity: 49.7 pg/mL
Standard: 10000 pg/mL
Detection Range: 156.25-10000 pg/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Metabolic pathway;Nutrition metabolism;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mouse CNCbl. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse CNCbl. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mouse CNCbl, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mouse CNCbl in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
10000.00 1.941 1.847
5000.00 1.625 1.531
2500.00 1.162 1.068
1250.00 0.921 0.827
625.00 0.453 0.359
312.50 0.366 0.272
156.25 0.235 0.141
0.00 0.094 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant CNCbl and the recovery rates were calculated by comparing the measured value to the expected amount of CNCbl in samples.
Matrix Recovery range Average
serum(n=5) 87-97% 90%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 81-97% 89%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of CNCbl and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 83-99% 98-105% 96-101% 88-95%
EDTA plasma(n=5) 98-107% 90-99% 78-96% 86-92%
Heparin plasma(n=5) 96-105% 89-103% 85-94% 87-96%
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