DESCRIPTION

Neural cell adhesion molecule 1 (NCAM1, also known as CD56) is a cell adhesion glycoprotein of the immunoglobulin (Ig) superfamily. It is a multifunction protein involved in synaptic plasticity, neurodevelopment, and neurogenesis. NCAM1 is expressed on human neurons, glial cells, skeletal muscle cells, NK cells and a subset of T cells, and the expression is observed in a wide variety of human tumors, including myeloma, myeloid leukemia, neuroendocrine tumors, Wilms' tumor, neuroblastoma, and NK/T cell lymphomas. Three major isoforms of NCAM1, with molecular masses of 120, 140, and 180 kDa, are generated by alternative splicing of mRNA. The glycosylphosphatidylinositol (GPI)-anchored NCAM120 and the transmembrane NCAM140 and NCAM180 consist of five Ig-like domains and two fibronection-type III repeats (FNIII). All three forms can be posttranslationally modified by addition of polysialic acid (PSA). Several other isofroms have also been described.

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	5.20	0.07	1.35
2	8	1.31	0.04	3.05
3	8	0.64	0.02	3.13

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	5.21	0.13	2.50
2	16	1.30	0.03	2.31
3	16	0.64	0.02	3.13

RECOVERY

The recovery of human NCAM1/CD56 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:800	99	95-104
	1:1,600	103	100-108
CSF	1:160	102	98-108

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 plasma was initially diluted 1:100. The human cerebrospinal fluid (CSF) was initially diluted 1:10.)

		Human plasma	Human cerebrospinal fluid(CSF)
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	89	95
	Range (%)	84-94	95-96
1:8	Average% of Expected	87	93
	Range (%)	83-91	90-96
1:16	Average% of Expected	88	93
	Range (%)	84-91	83-103

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human NCAM1/CD56 One-Step ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

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