Recombinant Human Complement Component C1r Protein, CF
Recombinant Human Complement Component C1r Protein, CF Summary
Product Specifications
Met1-Asp705, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
1807-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 50 mM Tris and 150 mM NaCl, pH 7.5. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 50 mM Tris, pH 7.5
- Recombinant Human Complement Component C1r (rhC1r) (Catalog # 1807-SE)
- Substrate: Z-Gly-Arg-SBzl (MP Biomedicals, Catalog # SB007), 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 rhC1r to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer with 200 µM of DTNB.
- Load 50 µL of the diluted rhC1r into a clear plate, and start the reaction by adding 50 µL of the Substrate/DTNB mixture to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate Mixture without any rhC1r.
- Read in kinetic mode for 5 minutes at an 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:
- rhC1r: 0.100 µg
- DTNB: 100 µM
- Substrate: 100 µM
Reconstitution Calculator
Background: Complement Component C1r
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 1-705) of human C1r was expressed, which had the Leu152 natural variant (3). The purified protein corresponded to the processed active form, with A and B chains starting at residue Ser18 and Ile464, respectively.
- Arlaud, G.J. et al. (2002) Biochem. Soc. Trans. 30:1001.
- Lacroix, M. et al. (2001) J. Biol. Chem. 276:36233.
- Journet A. and M. Tosi (1986) Biochem. J. 240:783.
Citations for Recombinant Human Complement Component C1r 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.
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Citations: Showing 1 - 4
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C1r/C1s deficiency is insufficient to induce murine systemic lupus erythematosus
Authors: K Uchio-Yama, M Tanaka, N Manabe
Genes Immun., 2018-03-18;0(0):.
Applications: Western Blot -
Complement C1 esterase inhibitor levels linked to infections and contaminated heparin-associated adverse events.
Authors: Zhou ZH, Chen T, Arora K
PLoS ONE, 2012-04-13;7(4):e34978.
Species: Human
Sample Types: Recombinant Protein
Applications: Binding Assay -
Complement C3a, CpG oligos, and DNA/C3a complex stimulate IFN-alpha production in a receptor for advanced glycation end product-dependent manner.
Authors: Ruan BH, Li X, Winkler AR, Cunningham KM, Kuai J, Greco RM, Nocka KH, Fitz LJ, Wright JF, Pittman DD, Tan XY, Paulsen JE, Lin LL, Winkler DG
J. Immunol., 2010-09-03;185(7):4213-22.
Species: Human
Sample Types: Recombinant Protein
Applications: Binding Assay -
Unique phenotypes of C1s deficiency and abnormality caused by two compound heterozygosities in a Japanese family.
Authors: Abe K, Endo Y, Nakazawa N, Kanno K, Okubo M, Hoshino T, Fujita T
J. Immunol., 2009-02-01;182(3):1681-8.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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