DESCRIPTION

Basal cell adhesion molecule (Lu/BCAM) is a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that plays important roles in cell adhesion and signaling. It is expressed on various cell types including erythrocytes, where it defines the Lutheran and Auberger blood group antigens. BCAM, involved in cell adhesion and migration, can also promote tumor metastasis and has been elucidated to play a functional role in the metastasis of thyroid cancer and gastric cancer. Research has linked BCAM to several pathological conditions.

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 (ng/mL)	SD	CV%
1	8	5.18	0.07	1.35
2	8	1.32	0.03	2.27
3	8	0.65	0.01	1.54

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	5.18	0.28	5.41
2	16	1.28	0.04	3.13
3	16	0.64	0.02	3.13

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Human plasma	1:16	79	74-81
Cell culture supernatant	1:2	88	83-96
	1:4	91	87-97

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.

		Human plasma	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	109	102
	Range (%)	104-113	102-103
1:8	Average% of Expected	111	118
	Range (%)	108-114	113-123
1:16	Average% of Expected	103	126
	Range (%)	88-117	124-129

CALIBRATION

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