Recombinant Human Complement Component C2 Protein, CF

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

Recombinant Human 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 >35 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Complement Component C2 protein
  Ala21-Leu752, with a C-terminal 6-His tag
• Accession #: P06681
• N-terminal Sequence: Ala21
• Structure / Form: Pro form
• 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: 82 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: 103 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 Tris, NaCl and CaCl₂.
• 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 Human Complement Component C2 (rhC2) (Catalog # 1936-SE)
  • Substrate: Z-Gly-Arg-SBzl
  • 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB)
  • Clear 96 well Plate
  • Plate Reader
  1. Dilute rhC2 to 4 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer with 200 µM of DTNB.
  3. Load 50 µL of the diluted rhC2 into a plate, and start the reaction by adding 50 µL of the Substrate/DTNB mixture to wells. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of the Substrate/DTNB mixture.
  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 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 0.32 cm
  Note: the output of many spectrophotometers is in mOD Per Well:
  • rhC2: 0.2 µg
  • DTNB: 100 µM
  • Substrate: 100 µM

Notes

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

Alternate Names for Recombinant Human 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

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 activation by C1, the single-chain form of C2 (amino acid residues 21‑752) becomes two chains, which are referred to as C2A and C2B. C2A (residues 244‑752) consists of a vWF domain (residues 254‑452) and a serine protease domain (residues 466‑752). C2B (residues 21‑243) contains 3 Sushi (SCR) domains. C2A, then combines with complement factor 4B to generate the C3 or C5 convertase. The full‑length of human C2 was expressed, and the purified protein corresponded to the single-chain form with the peptidase activity as described in the Activity Assay Protocol.

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

Publications for Complement Component C2 (1936-SE)(1)

Publication using 1936-SE

• Brudner , Matthew, Karpel , Marshall, Lear , Calli, Chen , Li, Yantosca , L Michae, Scully , Corinne, Sarraju , Ashish, Sokolovska , Anna, Zariffard , M Reza, Eisen , Damon P, Mungall , Bruce A, Kotton , Darrell, Omari , Amel, Huang , I-Chueh, Farzan , Michael, Takahashi , Kazue, Stuart , Lynda, Stahl , Gregory, Ezekowitz , Alan B, Spear , Gregory, Olinger , Gene G, Schmidt , Emmett V, Michelow , Ian C Lectin-dependent enhancement of Ebola virus infection via soluble and transmembrane C-type lectin receptors. PLoS ONE, 2013-04-02;8(4):e60838. 2013-04-02 [PMID: 23573288] (Bioassay, Human)

Bioinformatics

• Gene Symbol: C2
• Uniprot:
  • P06681()