Recombinant Mouse Complement Component C2 Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse Complement Component C2 Protein, CF Summary

• Details of Functionality: 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.
• Source: Mouse myeloma cell line, NS0-derived mouse Complement Component C2 protein
  Met1-Leu760, with a C-terminal 6-His tag.
• Accession #: O70350
• N-terminal Sequence: Leu20 & Lys251
• Protein/Peptide Type: Recombinant Enzymes
• Gene: C2
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 58 kDa and 84 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 66-75 kDa & 90-110 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • 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 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
  Note: the output of many spectrophotometers is in mOD Per Well:
  • rmC2: 0.200 μg
  • Substrate: 100 μM
  • DTNB: 100 μM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Complement Component C2 Protein, CF

• C2
• C3/C5 convertase
• CO2
• complement C2
• complement component 2
• Complement Component C2
• DKFZp779M0311
• EC 3.4.21
• EC 3.4.21.43

Background

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.

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

Publications for Complement Component C2 (6725-SE)(1)

Publication using 6725-SE

• T Zhou, Y Li, X Li, F Zeng, Y Rao, Y He, Y Wang, M Liu, D Li, Z Xu, X Zhou, S Du, F Niu, J Peng, X Mei, SJ Ji, Y Shu, W Lu, F Guo, T Wu, TF Yuan, Y Mao, B Peng Microglial debris is cleared by astrocytes via C4b-facilitated phagocytosis and degraded via RUBICON-dependent noncanonical autophagy in mice Nature Communications, 2022-10-24;13(1):6233. 2022-10-24 [PMID: 36280666] (Bioassay, Mouse)

Bioinformatics

• Gene Symbol: C2
• Uniprot:
  • O70350()