FAQs - Proteins & Antibodies
Here you find frequently asked questions about ProteaImmun’s proteins and antibodies.
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Do you know the epitope for anti-MT1-MMP pAb?
The polyclonal antibody against MT1-MMP was raised in rabbits against a peptide from the hinge region between catalytic and hemopexin domains of MT1-MMP.
Does MMP 13 catalytic domain (catalog number: 30100812) need any activation step with mercury containing compound (like APMA) before carrying out incubation with fluoregenic probe?
No, you don’t need to activate MMP-13 catalytic domain prior reaction with the fluorescent peptide substrate. The buffer you need is named Peptide hydrolysis buffer: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 0.025 % Brij 35.
Can you briefly describe the activation reaction for MT1-MMP Prodomain-Catalytic domain?
The activation of MT1-MMP Prodomain-Catalytic domain take place at 25 °C for 18 min in the following buffer: 50 mM Tris pH7.5 (HCl), 100 mM NaCl, 5 mM CaCl2 and 0.025 % Brij35. The reactants have the concentration of 0.05 µg/ml for MT1-MMP Pro-Cat and 0.5 µg/ml for Trypsin. The reaction is stopped with 10 µg/ml Aprotinin.
Can recombinant proteins be concentrated up to 0.2 mg/ml?
No, in this case it will not work. If concentration is increased the stability of the protein starts to decrease and it begins self-cleavage.
How do the lots quality is controlled?
Standard quality control for each new lot is performed by measuring protein concentration (BCA kit by Pierce) and SDS-PAGE / Coomassie staining, as described in the datasheet. Furthermore, we test cleavage by ADAMTS4 since we also use aggrecan-IGD as a substrate in our ELISA kits for determining aggrecanase activity. However, this is not part of the standard procedure, but however additional quality control.
Are your recombinant proteins tested for sterility and it is appropriate to use in tissue culture procedures?
Our recombinant proteins are not tested for sterility and therefore you should not use it for tissue culture.