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Active Tissue Inhibitors Of Metalloproteinase 3 (TIMP3)

SFD; TIMP Metallopeptidase Inhibitor 3; Metallopeptidase Inhibitor 3; Sorsby Fundus Dystrophy; Pseudoinflammatory

ACTIVITY TEST

Tissue Inhibitors of Metalloproteinase 3 (TIMP3) is a key extracellular matrix protein encoded by the TIMP3 gene, playing pivotal roles in regulating matrix metalloproteinase (MMP) activity. It is widely expressed in various tissues, participating in tissue remodeling, angiogenesis, and inflammation. Dysregulation of TIMP3 is linked to multiple diseases, including cancer, cardiovascular disorders, and degenerative conditions, due to its ability to inhibit MMP-mediated extracellular matrix degradation. TIMP3 binds specifically to MMP9 with high affinity, forming a stable complex that potently suppresses MMP9’s proteolytic activity to maintain tissue homeostasis.Thus a functional ELISA assay was conducted to detect the interaction of recombinant cattle TIMP3 and recombinant human MMP9. Briefly, TIMP3 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to MMP9-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-TIMP3 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant cattle TIMP3 and recombinant human MMP9 was shown in Figure 1, the EC50 for this effect is 0.83911µg/mL.

USAGE

Reconstitute in 100mM NaHCO3, 500mM NaCl (pH8.3) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Neuroimmunomodulation Upregulated Expression of Matrix Metalloproteinases and Tissue Inhibitors of Matrix Metalloproteinases in BALB/c Mouse Brain Challenged with Japanese Encephalitis Virus Pubmed: source
Experimental Biology and Medicine Acidic pH conditions mimicking degenerative intervertebral discs impair the survival and biological behavior of human adipose-derived mesenchymal stem cells Rsmjournals: Source
PLoS ONE A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic Approaches Plosone: Source
Physiol. Res. The unwounded skin remodeling in animal models of diabetes types 1 and 2. Pubmed: 24020818
VirusDisease Circulating levels of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases during Japanese encephalitis virus infection Pubmed:26925446
Acta Pharmacol Sin. Pharmacological evidence: a new therapeutic approach to the treatment of chronic heart failure through SUR2B/Kir6.1 channel in endothelial cells. pubmed:27890915
Biochemical genetics Comparison of Selected Protein Levels in Tumour and Surgical Margin in a Group of Patients with Oral Cavity Cancer pubmed:28421310
International Journal of Cancer Circulating and tissue IMP3 levels are correlated with poor survival in renal cell carcinoma Pubmed: 30650187
J. Pers. Med Altered TIMP-3 Levels in the Cerebrospinal Fluid and Plasma of Patients with Alzheimer's Disease Pubmed:35629249
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