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Recombinant Human Complement MASP3 Catalytic Domain, CF

Catalog #: 1724-SE
1 Citation Datasheet / COA / SDS
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1724-SE-020
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Recombinant Human Complement MASP3 Catalytic Domain, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
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). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468. The specific activity is >10,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Complement Factor MASP3 protein
Ile450-Val721, with an N-terminal signal peptide and a C-terminal 10-His tag
Accession #
Q96RS4
N-terminal Sequence
Analysis
Ile450
Predicted Molecular Mass
31 kDa
SDS-PAGE
43 kDa, reducing conditions

Product Datasheets

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1724-SE

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1724-SE

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, pH 8.5
  • Recombinant Human Complement Factor MASP3 Catalytic Domain (rhMASP3) (Catalog # 1724-SE)
  • Substrate: Z-Lys-SBzl (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
  • UV-transparent 96 Well Microplate (Corning, Catalog # 3635)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhMASP3 to 0.04 ng/µL in Assay Buffer.
  2. Combine equal volumes of 10 mM Substrate and 10 mM DTNB for 5 mM of each.
  3. Dilute Substrate/DTNB mixture to 200 µM of each with Assay Buffer.
  4. Load 50 µL of the 0.04 ng/µL rhMASP3 into a UV plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 μL of 200 µM Substrate without any rhMASP3.
  5. Read at a wavelength of 405 nm (bottom read) in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 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
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhMASP3: 0.002 µg
  • Substrate: 100 µM
  • DTNB: 100 µM
Reconstitution Calculator

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Background: Complement Factor MASP3

MASP3 is a member of the MASPs involved in mannan-binding lectin (MBL) complement pathway (1). The MBL pathway is initiated by the binding of MBL to specific carbohydrate structures found on the surface of a variety of microorganisms. Activation of the complement pathway via MBL is initiated by specific MASPs. Three MASPs have been identified and all have domain structures similar to those of C1r and C1s with a heavy chain (chain A) and a light chain (chain B). Chain A is composed of CUB1, EGF, CUB2, CCP1 and CCP2 while chain B corresponds to the catalytic domain found in many serine proteases. MASP1 and MASP3 are two alternatively spliced products of a single gene, which contain the same A chains but entirely different B chains. Distinct MASPs found in different MBL oligomers may have different biological activities. For example, MASP3, found together with MASP2, downregulates the C4 and C2 cleaving activity of MASP2. The protease activity of MASP3 is first revealed here using recombinant human MASP3CD (2), which is inhibited by serine protease inhibitors such as ecotin and AEBSF (Catalog # 1328-PI and EI001).

References
  1. Dahl, M.R. et al. (2001) Immunity 15:127.
  2. Cortesio, C.L. and W. Jiang (2006) Arch. Biochem. Biophys. 449:164.
Long Name
Mannan-binding lectin-Associated Serine Protease 3
Entrez Gene IDs
5648 (Human)
Alternate Names
Complement Factor MASP3; Complement factor MASP-3; Complement-activating component of Ra-reactive factor; CRARF1; CRARFMAP1; DKFZp686I01199; EC 3.4.21; EC 3.4.21.-; EC 3.4.21.104; EC 3.4.21.42; FLJ26383; mannan-binding lectin serine peptidase 1 (C4/C2 activating component ofRa-reactive factor); mannan-binding lectin serine protease 1 (C4/C2 activating component ofRa-reactive factor); mannan-binding lectin serine protease 1; Mannose-binding lectin-associated serine protease 1; Mannose-binding protein-associated serine protease; MAp44; MASP; MASP1; MASP-1; MASP3; MGC126283; MGC126284; PRSS5; Ra-reactive factor serine protease p100; RaRF; Serine protease 5

Citation for Recombinant Human Complement MASP3 Catalytic Domain, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Mannan-binding lectin-associated serine protease 3 cleaves synthetic peptides and insulin-like growth factor-binding protein 5.
    Authors: Cortesio CL, Jiang W
    Arch. Biochem. Biophys., 2006-03-03;449(1):164-70.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay

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