DESCRIPTION

MMP9 (matrix metallopeptidase 9), also named as Gelatinase B, is a member of matrix metalloproteinase (MMP) family. The MMP family of enzymes is comprised of critically important extracellular matrix remodeling proteases whose activity has been implicated in normal embryogenesis, tissue remodelling and many diseases such as arthritis, cancer, periodontitis, glomerulonephritis, encephalomyelitis, atherosclerosis and tissue ulceration. These proteases have come to represent important therapeutic and diagnostic targets for the treatment and detection of human cancers. MMP9 is produced by a variety of normal and transformed cells including neutrophils, monocytes, macrophages, astrocytes, fbroblasts, osteoclasts and so on. In tumors, MMP-9 destroys collagen (type IV) in the vascular basal membrane in the vicinity of tumor cells which invade the surrounding tissues and contributes to metastasis. Circulating levels of MMP-9 are increased in many inflammatory disorders such as atherosclerosis, hepatitis C virus infection and colorectal cancer.

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 (pg/mL)	SD	CV%
1	8	1,259.7	74.6	5.9
2	8	321.8	13.7	4.2
3	8	165.1	2.3	1.4

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,247.6	77.9	6.2
2	16	312.1	17.1	5.5
3	16	161.1	6.4	4.0

RECOVERY

The recovery of human MMP-9 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:1,600	96	92-98
Cell culture supernatant	1:1,600	92	87-95
Urine	1:4	99	98-101
	1:8	93	87-96
Saliva	1:800	100	97-107

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 1:100. The cell culture supernatant was initially diluted 1:100. The urine was initially diluted 1:2. The saliva was initially diluted 1:50.)

		Human serum	Cell culturesupernatant	Urine	Saliva
1:2	Average% of Expected	100	100	100	100
	Range (%)	-	-	-	-
1:4	Average% of Expected	104	100	113	99
	Range (%)	98-107	99-101	109-117	97-101
1:8	Average% of Expected	106	97	121	102
	Range (%)	98-113	94-98	117-125	99-106
1:16	Average% of Expected	109	97	122	100
	Range (%)	95-126	94-100	118-125	95-108

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human MMP-9 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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• Price: USD, per kit (48T / 96T options available)