Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human MMP-12 (carrier-free)

764204

25 μg

411 USD

BA

764204

Recombinant Human MMP-12 (carrier-free)

Macrophage metalloelastase (MME), Matrix metalloproteinase 12

25 μg

411 USD

Human

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RECOM

Human MMP-12 Activity assay; Human MMP-12 (hMMP-12) activity is measured by its ability to cleave a fluorogenic peptide substrate Mca-PLGL(Dpa)AR after its activation in the presence of p-Aminophenylmercuric acid. The increase of the product is monitored by increase in intensity of fluorescence at 405 nm with excitation at 320 nm. This protein is in the latent form and needs to be activated for bioassay.; Materials;Assay Buffer: TCNB (50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35, pH 7.5);Recombinant Human MMP-12;MCa-PLGL(Dpa)AR (AnaSpec Cat # AS27076; 2 mM in DMSO);P-Aminophenylmercuric acid (Calbiochem Cat# 164610; 100 mM in DMSO); Activity assay procedures;Dilute hMMP-12 in the assay buffer at 100 µg/mL.;Activate hMMP-12 by adding APMA to a final concentration of 1 mM.;Incubate the activating mixture for 1 hr at 37 °C.;Dilute the activated hMMP-12 to 0.4 ug/mL (0.4 ng/µL) in Assay Buffer.;Dilute the substrate at 20 µM in Assay Buffer.;Load into a well plate 50 μL of the 0.4 ng/μL hMM-12 and start the reaction by adding 50 μL of 20 μM Substrate. Include a substrate blank containing 50 μL Assay Buffer and 50 μL of 20 μM Substrate without any hMMP-12.;Read the product formation by measuring 320/ 405 nm (Excitation/Emission) in kinetic mode for 5 minutes.;The final hMMP-12 concentration is 0.2 µg/mL (0.2 ng/µl, 20 ng);The final concentration of the substrate is 10 µM.;BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.;