DESCRIPTION

CD147, also called Basigin (BSG) or EMMPRIN, is a transmembrane glycoprotein that belongs to the immunoglobulin superfamily and is important in numerous physiological and pathological phenomena. It is involved in reproduction, neural function, inflammation and tumor invasion. CD147 is broadly distributed, and expressed at particularly high levels at the retinal pigment epithelium and neonatal blood-brain barrier, on tumour cells and on activated T cells. CD147 can stimulate adjacent fibroblasts to produce matrix metalloproteinases (MMPS). It has been considered as an objective and effective marker to predict invasion and prognosis in various cancers. Soluble CD147 (sCD147) has been uncovered in addition to membrane-bound CD147. sCD147 production is mediated by two different mechanisms: microvesicular secretion and proteolytic cleavage. The presence of sCD147 in body fluids is associated with several diseases such as systemic sclerosis, hepatocellular carcinoma, ankylosing spondylitis and coronary diseases.

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 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	20	1,022.6	61.6	6.0
2	20	223.5	6.9	3.1
3	20	59.1	2.3	3.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	1,060.9	67.6	6.4
2	24	239.5	13.6	5.7
3	24	70.1	5.2	7.5

RECOVERY

The recovery of human CD147 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:16	103	98-111
	1:32	99	88-117
Cell culture supernatant	1:4	99	87-111
	1:8	102	89-113
Urine	1:20	111	100-119
	1:40	101	73-119
Saliva	1:8	90	75-100
	1:16	88	76-112
Human milk	1:16	101	94-110
	1:32	110	100-123

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 1:2. The urine was initially diluted 1:2.5. The human milk was initially diluted 1:2.)

		Human serum(Sample DiluentPT 1-ef)	Cell culturesupernatant (SampleDiluent PT 4)	Urine (SampleDiluent PT 1-ef)	Saliva (SampleDiluent PT 1-ef)	Human milk(Sample DiluentPT 4B1)
1:2	Average% ofExpected	100	100	100	100	100
	Range (%)	-	-	-	-	-
1:4	Average% ofExpected	95	115	117	106	103
	Range (%)	89-100	107-123	112-121	96-116	98-110
1:8	Average% ofExpected	103	94	102	89	88
	Range (%)	86-114	88-101	101-104	87-92	85-90
1:16	Average% ofExpected	82	-	75	115	74
	Range (%)	70-90	-	72-78	110-119	72-75

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human CD147 produced at Sunlong Medical™.

Ordering & shipping

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