Assay Principle

This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture FBLN5 from samples and standards. After washing, biotin-labeled Detection Antibody is added and incubated, followed by washing to form a "Capture Antibody-Antigen-Detection Antibody" immune-complex. Subsequently, Streptavidin-Horseradish Peroxidase (SA-HRP) is added and incubated. After incubation and washing, TMB Substrate is added for color development. If the target analyte is present in the sample, a blue color develops. Stop Solution is then added to terminate the reaction. During the assay, unbound components are washed away. The Optical Density (OD) is measured at 450 nm using a microplate reader. The intensity of the color is proportional to the FBLN5 concentration in the sample, and the concentration is calculated by plotting a standard curve.

Precision

Intra-assay and inter-assay Coefficients of Variation (CV) are both <10%.

· Intra-assay Precision: Three known concentration samples were assayed 20 times on one plate. The CV of the concentrations was calculated.

· Inter-assay Precision: Three known concentration samples were assayed in 20 replicates across three different plates. The CV of the concentrations was calculated.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	0.29	1.4	2.8	0.31	1.8	3.3
Standard Deviation	0.004	0.013	0.035	0.006	0.021	0.047
CV (%)	1.38	0.93	1.25	1.94	1.17	1.42

Recovery

Recovery was tested by spiking known concentrations of Human FBLN5 into different sample matrices. The recovery range and average recovery are shown below.

 

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	89-101	96
Plasma (n=8)	91-104	98
Cultured Cells (n=8)	100-117	107

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