DESCRIPTION

TIMP2, a 21-kDa protein, is a member of the family of matrix metalloproteinase inhibitors, which contains four members (TIMP1, TIMP2, TIMP3, and TIMP4). Tissue inhibitors of metalloproteinases (TIMPs) are multifaceted molecules that exhibit properties beyond their classical proteinase inhibitory function. TIMP-2 is the only TIMP functioning as both an MMP inhibitor and an activator, because it can interact with membrane type 1 MMP and pro-MMP2 to facilitate the activation of pro-MMP2 to its active form. TIMP2 can regulate many physiological progression such as tumor growth, metastasis, and angiogenesis through matrix metalloproteinases (MMPs). TIMP2 knockout resulted in constitutive MMP9 activation abundantly and adverse MI remodeling in vivo.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with biotin) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with biotin also binds to the analyte. Streptavidin-HRP binds to the biotin. 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	987.5	40.1	4.1
2	20	255.8	7.9	3.1
3	20	84.9	3.2	3.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	971.0	31.5	3.2
2	24	262.8	7.5	2.8
3	24	88.7	2.4	2.7

RECOVERY

The recovery of human TIMP2 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:1,500	93	72-119
	1:3,000	105	93-113
Cell culture supernatant	1:1,200	95	90-99
	1:2,400	99	90-116
Urine	1:15	108	104-111
	1:30	106	98-117
Saliva	1:80	106	102-113
	1:160	100	94-106

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 samples were initially diluted 1:200. The cell culture supernatant samples were initially diluted 1:160. The urine samples were initially diluted 1:2. The saliva samples were initially diluted 1:10. )

		Human serum	Cell culturesupernatant	Urine	Saliva
1:2	Average% of Expected	100	100	100	100
	Range (%)	-	-	-	-
1:4	Average% of Expected	99	101	105	103
	Range (%)	97-101	100-101	93-124	99-108
1:8	Average% of Expected	101	101	108	103
	Range (%)	92-106	100-101	93-123	100-108
1:16	Average% of Expected	100	99	114	102
	Range (%)	95-105	93-105	98-125	94-107

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human TIMP2 produced at Sunlong Medical™.

Ordering & shipping

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