Recombinant Human Complement Component C2 Protein, CF

R&D Systems | Catalog # 1936-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Complement Component C2 Protein (1936-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

P06681

Structure / Form

Pro form

Applications

Enzyme Activity
View all Complement Component C2 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Complement Component C2 protein
Ala21-Leu752, 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

Ala21

Predicted Molecular Mass

82 kDa

SDS-PAGE

103 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 >35 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

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.

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.

Alternate Names

C2

Entrez Gene IDs

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

Gene Symbol

C2

UniProt

P06681

Additional Complement Component C2 Products

Product Documents for Recombinant Human Complement Component C2 Protein, CF

Certificate of Analysis

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

For research use only

Citations for Recombinant Human Complement Component C2 Protein, CF

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Protocols

View specific protocols for Recombinant Human Complement Component C2 Protein, CF (1936-SE):

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

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