Buffer Formulation	20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300.
Traits	Freeze-dried powder
Purity	> 97%
Isoelectric Point	5.1
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Matrix Metalloproteinase 13 (MMP13) is a member of the matrix metalloproteinase (MMP) family. MMP13 has been proposed to participate in aggrecan degradation associated with osteoarthritis and cleavage of type II collagen in osteoarthritic cartilage explants and in tumor progression and metastasis. In addition, it can cleave type I, III, IV, IX, X and XIV collagens and fibronectin. MMP13 is likely to play a crucial role in the modulation of extracellular matrix degradation and cell-matrix interactions. Although gelatin zymography is mainly used for the detection of the MMP2 and MMP9, it also can be used for MMP13 dection. Briefly, various concentrations of MMP13 (1000ng, 500ng, 250ng, 125ng) were denatured by SDS loading buffer, electrophoresed through sodium dodecylsulphat- polyacrylamide gel (SDS-PAGE; 10% gels) containing gelatin (1mg/mL) with nonreducing conditions. After renaturation, incubation and CCB-stained, active MMP13 would hydrolyze gelatin nearby, which was indicated by the white bands on the gel. In this experiment we use heat-denatured MMP13 protein as negative control, and blood sample as positive control. Result: Gelatin hydrolysis by recombinant rat MMP13 was shown in figure 1.

 

USAGE

 

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) 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.

 

Image


SDS-PAGE Image

 

Figure. Western Blot; Sample: Recombinant MMP13, Rat.

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