DESCRIPTION

CEACAM6, also known as CD66c, is a glycosylphosphatidylinositol (GPI)-linked cell surface protein that belongs to the CEACAM family of the immunoglobulin superfamily. It is normally expressed on the surface of myeloid and epithelial surfaces and upregulated preferentially at the apical/luminal membranes of many tumors. CEACAM6 plays an important role in cell adhesion, proliferation, tumorigenesis, metastasis, and immunity.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	260.6	17.3	6.6
2	8	66.8	1.1	1.6
3	8	35.7	0.8	2.2

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	255.6	13.3	5.2
2	16	66.7	1.4	2.1
3	16	35.5	0.8	2.3

RECOVERY

The recovery of human CEACAM6 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:800	99	91-105
Cell culture supernatant	1:2	113	105-120

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The human serum was initially diluted 1:50.)

		Human serum	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	96	108
	Range (%)	82-110	98-118
1:8	Average% of Expected	86	102
	Range (%)	80-92	99-105
1:16	Average% of Expected	84	97
	Range (%)	73-96	96-98

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human CEACAM6 produced at Sunlong Medical™.