Babco, Signet, Sternberger Monoclonals, Senetek, Covance

protein

Recombinant Human Complement Factor D (carrier-free)

791802

10 μg

218 USD

BA

791802

Recombinant Human Complement Factor D (carrier-free)

DF, ADN, PFD, Adipsin, Properdin Factor D, C3 Convertase Activator, Complement Factor D (Adipsin)

10 μg

218 USD

Human

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RECOM

Assay ProcedureRecombinant Human complement Factor D/Adipsin activity is measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Lys-Thiobenzyl ester (Z-Lys-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). The increase of the product is monitored by increase in intensity of absorbance at 405 nm.Materials;Assay buffer: 50 mM Tris, 1 M NaCl, pH 7.5;Recombinant human Complement Factor D/Adipsin;Substrate: Z-Lys-SBzl (Bachem, Cat. No. M-1300), 10 mM stock in DMSO;5,5’Dithio-bis-(2-nitrobenzoic acid) (DTNB) (Sigma, Cat. No. D-8130), 10 mM stock in DMSO;96-well clear plate (Costar, Cat. No. 92592);Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent;Procedure;Dilute the recombinant factor D to 5 ng/µL in assay buffer.;Dilute the substrate to 200 µM in assay buffer with 200 µM DTNB.;Load 50 µL of the diluted recombinant factor D into a clear plate, and start the reaction by adding 50 µL of substrate/DTNB mixture to the wells. Include a substrate blank containing 50 µL assay buffer and 50 µL substrate mixture without any recombinant Factor D.;Read in kinetic mode for 5 minutes at an absorbance of 405 nm.;Calculate specific activity:;Specific Activity (pmol/min/µg) = Adjusted Vmax* (OD/min) x well volume (L) x 10^12 pmol/mol ext. coeff** (M^-1cm^-1) x path corr.*** (cm) x amount of enzyme (µg);*Adjusted for substrate blank**Using the extinction coefficient 13260 M-1cm-1***Using the path correction 0.320 cm;Recombinant factor D: 0.25 µg;DTNB: 100 µM;Substrate: 100 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.;