DESCRIPTION

L1CAM, also known as NCAM-L1 or CD171, is a cell adhesion molecule of the immunoglobulin superfamily. It is a 200-220 kDa transmembrane glycoprotein composed of six Ig-like domains and five fibronectin type III repeats followed by a transmembrane region and a highly conserved cytoplasmic tail. L1CAM is primarily expressed in the nervous system and is involved in neuronneuron adhesion, neurite fasciculation, outgrowth of neurites, cerebellar granule cell migration, neurite outgrowth on Schwann cells and interactions among epithelial cells of intestinal crypts. L1CAM is overexpressed in many human cancers and is often associated with bad prognosis.

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	10.11	0.13	1.29
2	8	2.60	0.03	1.15
3	8	1.35	0.02	1.48

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	9.94	0.22	2.21
2	16	2.56	0.05	1.95
3	16	1.33	0.03	2.26

RECOVERY

The recovery of mouse L1CAM spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:256	107	104-113
	1:512	115	115-116
Tissue lysate	1:20	104	101-106
	1:40	110	107-114

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 mouse serum was initially diluted 1:32. The tissue lysate was initially diluted 1:40.)

		Mouse serum	Tissue lysate
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	98	112
	Range (%)	92-103	111-113
1:8	Average% of Expected	98	106
	Range (%)	92-104	105-108
1:16	Average% of Expected	98	109
	Range (%)	92-105	93-125

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant mouse L1CAM produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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