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Recombinant Human Complement MASP3 Catalytic Domain, CF

R&D Systems | Catalog # 1724-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Complement MASP3 Catalytic Domain (1724-SE)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

Q96RS4

Applications

Enzyme Activity
View all Complement Factor MASP3 Proteins and Enzymes »

Product Specifications

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

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.

N-terminal Sequence Analysis

Ile450

Predicted Molecular Mass

31 kDa

SDS-PAGE

43 kDa, reducing conditions

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.

Formulation, Preparation, and Storage

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.

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 # http://www.rndsystems.com/product_results.aspx?k=1328-PI">1328-PI and http://www.rndsystems.com/product_results.aspx?k=EI001">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

Alternate Names

MASP1

Entrez Gene IDs

5648 (Human)

Gene Symbol

MASP1

UniProt

Q96RS4

Additional Complement Factor MASP3 Products

Product Documents for Recombinant Human Complement MASP3 Catalytic Domain, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Complement MASP3 Catalytic Domain, CF

For research use only

Citations for Recombinant Human Complement MASP3 Catalytic Domain, CF

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Protocols

View specific protocols for Recombinant Human Complement MASP3 Catalytic Domain, CF (1724-SE):

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

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