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Recombinant Human Complement Component C2 Protein, CF

Catalog #: 1936-SE
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
1936-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Complement Component C2 Protein, CF Summary

Product Specifications

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.
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 >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
Analysis
Ala21
Structure / Form
Pro form
Predicted Molecular Mass
82 kDa
SDS-PAGE
103 kDa, reducing conditions

Product Datasheets

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1936-SE

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1936-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.

Assay Procedure

Materials
  • 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 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:
  • rhC2: 0.2 µg
  • DTNB: 100 µM
  • Substrate: 100 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Complement Component C2

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.

References
  1. Arlaud, G.J. et al. (2002) Biochem. Soc. Trans. 30:1001.
Entrez Gene IDs
717 (Human); 12263 (Mouse); 24231 (Rat)
Alternate Names
C2; C3/C5 convertase; CO2; complement C2; complement component 2; Complement Component C2; DKFZp779M0311; EC 3.4.21; EC 3.4.21.43

Citation for Recombinant Human Complement Component C2 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Lectin-dependent enhancement of Ebola virus infection via soluble and transmembrane C-type lectin receptors.
    Authors: 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
    PLoS ONE, 2013;8(4):e60838.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay

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