Recombinant Mouse Complement Component C1ra Protein, CF

R&D Systems | Catalog # 7160-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Complement Component C1ra Protein (7160-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

AAH04637

Applications

Enzyme Activity
View all Complement Component C1ra Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Complement Component C1ra protein
Met1-Asn707, with a C-terminal 6-His tag

Purity

>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser17 and Ile463

Predicted Molecular Mass

51 kDa (heavy chain) & 28 kDa (light chain)

SDS-PAGE

58-59 kDa and 35-36 kDa, reducing conditions

Activity

Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-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 >5,500 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7160-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl, NaCl, and Glycerol.
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 Component C1ra

The classical complement pathway plays a major role in innate immunity against infection. This pathway is triggered by C1, a multimolecular complex composed of the recognition protein C1q and two serine proteases, C1r and C1s. Following the C1q recognition, C1r is autoactivated, and in turn activates C1s, which cleaves C4 and C2, the C1 substrates (1). Both C1r and C1s activation involve cleavage of a specific Arg-Ile bond, converting the single-chain proenzymes into active proteases composed of disulfide-linked heavy and light chains (2). The heavy chain contains multiple domains in the order of CUB1-EGF-CUB2-CCP1-CCP2-Activation Peptide. The light chain contains the serine protease catalytic domain. The mouse C1r gene is duplicated as C1rA and C1rB. The C1rA gene is primarily expressed in liver and is therefore the homologue of human C1r gene (3). The full-length mouse C1r-A was expressed and purified from conditioned medium. The purified recombinant mC1rA protein corresponds to the processed active form, with the A and B chains beginning at residues Ser 17 and Ile 463, respectively.

References

  1. Arlaud, G. J. et al. (2002) Biochem. Soc. Trans. 30:1001.
  2. Lacroix, M. et al. (2001) J. Biol. Chem. 276:36233.
  3. Garnier, G. et al. (2003) J. Biochem. 371:630.

Alternate Names

C1ra

Entrez Gene IDs

50909 (Mouse)

Gene Symbol

C1RA

UniProt

AAH04637

Additional Complement Component C1ra Products

Product Documents for Recombinant Mouse Complement Component C1ra Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Complement Component C1ra Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

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Protocols

View specific protocols for Recombinant Mouse Complement Component C1ra Protein, CF (7160-SE):

Materials
  • Assay Buffer: 50 mM Tris, pH 7.5
  • Recombinant Mouse Complement Component C1Ra (rmC1Ra) (Catalog # 7160-SE)
  • Substrate: Z-Gly-Arg-SBzl (MP Biomedicals, Catalog # SB007), 10 mM stock in DMSO
  • 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
  • 96 well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmC1Ra to 0.5 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer with 200 µM of DTNB.
  3. Load 50 µL of the diluted rhC1r into a clear plate, and start the reaction by adding 50 µL of the Substrate/DTNB mixture to wells.  Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate Mixture without any rmC1Ra.
  4. Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
  5. 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 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rmC1Ra: 0.025 μg
  • DTNB: 100 µM
  • Substrate: 100 µM

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