Recombinant Mouse Complement Component C2 Protein, CF

R&D Systems | Catalog # 6725-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Complement Component C2 Protein (6725-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

O70350

Applications

Enzyme Activity
View all Complement Component C2 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Complement Component C2 protein
Met1-Leu760, with a C-terminal 6-His tag.

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Leu20 & Lys251

Predicted Molecular Mass

58 kDa and 84 kDa

SDS-PAGE

66-75 kDa & 90-110 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 >100 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6725-SE
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
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 C2

Complement Component C2 (C2) is a serine protease that is part of the classical pathway of the complement system. 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 (1). After proteolytic activation by C1, the single chain form of C2 (amino acid residues 19‑760) becomes two chains, which are referred to as C2A and C2B. C2A (residues 251‑760) consists of a vWF domain (residues 260‑457) and a serine protease domain (residues 480‑760). After the activation by C1, C2A combines with complement factor 4B to generate the C3 or C5 convertase. The full length mouse C2 was expressed, and the purified protein consists of a mixture of single chain and C2A form based on the N-terminal sequencing data.

References

  1. Arlaud, G.J. et al. (2002) Biochem. Soc. Trans. 30:1001.

Alternate Names

C2

Entrez Gene IDs

717 (Human); 12263 (Mouse); 24231 (Rat)

Gene Symbol

C2

UniProt

O70350

Additional Complement Component C2 Products

Product Documents for Recombinant Mouse Complement Component C2 Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse Complement Component C2 Protein, CF

For research use only

Citations for Recombinant Mouse Complement Component C2 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Complement Component C2 Protein, CF (6725-SE):

Materials
  • Assay Buffer: 50 mM Tris, pH 8.0
  • Recombinant Mouse Complement Component C2 (rmC2) (Catalog # 6725-SE)
  • Substrate: Z-GR-SBzl, 10 mM stock in DMSO
  • 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), 10 mM stock in DMSO
  • Clear 96-well Plate
  • Plate Reader
  1. Dilute rmC2 to 4 ng/μL in Assay Buffer.
  2. Dilute Substrate to 200 μM with 200 μM DTNB in Assay Buffer.
  3. Load into a plate 50 μL of 4 ng/μL rmC2, and start the reaction by adding 50 μL of 200 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 200 μM Substrate.
  4. Read plate at a wavelength of 405 nm (bottom read) in kinetic mode for 5 minutes.
  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 0.320 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rmC2: 0.200 μg
  • Substrate: 100 μM
  • DTNB: 100 μM

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