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Recombinant Mouse Complement Component C2 Protein, CF

Catalog #: 6725-SE
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
6725-SE-010
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Recombinant Mouse 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 >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
Analysis
Leu20 & Lys251
Predicted Molecular Mass
58 kDa and 84 kDa
SDS-PAGE
66-75 kDa & 90-110 kDa, reducing conditions

Product Datasheets

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6725-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.

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.

Assay Procedure

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
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

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.
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 Mouse 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. Microglial debris is cleared by astrocytes via C4b-facilitated phagocytosis and degraded via RUBICON-dependent noncanonical autophagy in mice
    Authors: 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
    Nature Communications, 2022-10-24;13(1):6233.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay

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