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. Following the C1q recognition, C1r is autoactivated, and in turn activates C1s, which cleaves C4 and C2, the C1 substrates (1). Both C1r and C1s activation involve cleavage of a specific Arg-Ile bond, converting single-chain proenzymes into active proteases of disulfide bond-linked chains (A and B) (2). The A chains contain multiple domains in the order of CUB1-EGF-CUB2-CCP1-CCP2-Activation Peptide. The B chains contain the serine protease catalytic domain. The full-length (amino acid residues 1-688) of human C1s was expressed (3-5). The purified protein corresponded to the processed active form, with A and B chains starting at residue Glu16 and Ile438, respectively.
Recombinant Human Complement Component C1s Protein, CF
R&D Systems | Catalog # 2060-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Complement Component C1s Protein (2060-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
Structure / Form
Disulfide-linked heterodimer
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Complement Component C1s protein
Met1-Asp688, with a C-terminal 6-His tag
Met1-Asp688, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Glu16 (A chain) & Ile438 (B chain)
Predicted Molecular Mass
47 kDa (A chain) & 28 kDa (B chain)
SDS-PAGE
62 kDa and 32 kDa, reducing conditions
Activity
Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Lys-ThioBenzyl ester (Z-Lys-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 >20,000 pmol/min/µg, as measured under the described conditions.
The specific activity is >20,000 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 2060-SE in the following applications:
Formulation, Preparation, and Storage
2060-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2 and Glycerol. |
| 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.
|
Background: Complement Component C1s
References
- Arlaud, G.J. et al. (2002) Biochem. Soc. Trans. 30:1001.
- Lacroix, M. et al. (2001) J. Biol. Chem. 276:36233.
- Tosi, M. et al. (1987) Biochemistry 26:8516.
- Mackinnon, C.M. et al. (1987) Eur. J. Biochem. 169:547.
- Kusumoto, H. et al. (1988) Proc. Natl. Acad. Sci. USA 85:7307.
Alternate Names
C1S
Entrez Gene IDs
716 (Human)
Gene Symbol
C1S
UniProt
Additional Complement Component C1s Products
Product Documents for Recombinant Human Complement Component C1s Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Complement Component C1s Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Complement Component C1s Protein, CF
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Protocols
View specific protocols for Recombinant Human Complement Component C1s Protein, CF (2060-SE):
Materials
*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:
- Assay Buffer: 50 mM Tris, 250 mM NaCl, pH 8.0
- Recombinant Human Complement Component C1s (rhC1s) (Catalog # 2060-SE)
- Substrate: N-Carbobenzyloxy-Lys-ThioBenzyl ester (Z-K-SBzl) (Bachem, Catalog # M-1300), 10 mM stock in DMSO
- 5,5’Dithio-bis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhC1s to 0.2 ng/μL in Assay Buffer.
- Dilute Substrate to 1000 μM containing 200 μM DTNB in Assay Buffer.
- Load in plate, 50 μL of 0.2 ng/μL rhC1s, and start the reaction by adding 50 μL of 1000 μM Substrate/200 μM DTNB mixture to wells. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 1000 μM Substrate/200 μM DTNB mixture.
- Read in kinetic mode for 5 minutes at absorbance of 405 nM.
- 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:
- rhC1s: 0.010 μg
- DTNB: 100 μM
- Substrate: 500 μM