Assay Principle

This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture TMPRSS6 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 TMPRSS6 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)	3.12	6.24	12.49	3.12	6.24	12.49
Standard Deviation	0.047	0.151	0.366	0.063	0.197	0.378
CV (%)	1.51	2.42	2.93	2	3.15	3.03

Recovery

Recovery was tested by spiking known concentrations of Human TMPRSS6 into different sample matrices. The recovery range and average recovery are shown below.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	90-103	96
Plasma (n=8)	93-102	99
Cultured Cells (n=8)	97-118	104

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