DESCRIPTION

CD43, also known as leukosialin, sialophorin, leukocyte sialoglycoprotein, SPN and W3/13 antigen, is a mucin-like type I transmembrane protein. It is also a sialoglycoprotein with an apparent molecular weight of ~100 kDa and is found on thymocytes, T lymphocytes, neutrophils, plasma cells and myelomas but not on B lymphocytes or most other tissues. CD43 is involved in the contact between cells participating in a series of events such as signaling, adherence, and host-parasite interactions.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns 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	17.60	0.56	3.18
2	8	4.35	0.11	2.53
3	8	1.63	0.10	6.13

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	16.87	0.96	5.69
2	16	4.20	0.22	5.24
3	16	1.70	0.11	6.47

RECOVERY

The recovery of mouse CD43/Leukosialin spiked to three different levels throughout the range of the assay in lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Lysate	1:128	95	80-124
	1:256	107	94-114

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 lysate was initially diluted 1:16.)

		Lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	105
	Range (%)	104-106
1:8	Average% of Expected	108
	Range (%)	107-109
1:16	Average% of Expected	103
	Range (%)	101-104

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse CD43/Leukosialin Sandwich ELISA Kit produced at Sunlong Medical™.