Recombinant Human Complement Component C1rLP His Protein, CF New
Recombinant Human Complement Component C1rLP His Protein, CF Summary
Product Specifications
Cys36-Asp487 with N-terminal HA (YPYDVPDYA) and 6-His tags
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.
11614-CP
| Formulation | Supplied as a 0.2 μm filtered solution in Tris 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.
|
Assay Procedure
- Assay Buffer: 50 mM Tris, pH 7.5
- Recombinant Human Complement Component C1rLP (rhC1rLP) (Catalog # 11614-CP)
- Substrate: Z-GR-SBzl,10 mM stock in DMSO
- 5,5'Dithio-bis (2-nitrobenzoic acid) (DTNB), 10 mM stock in DMSO
- Clear 96-well plate
- Plate Reader with Absorbance Read Capability
- Dilute rhC1rLP to 40 ng/µL in Assay Buffer.
- Prepare a Substrate Mixture containing 400 µM of Z-GR-SBzl and 200 µM of DTNB in Assay Buffer.
- In a plate, load 50 µL of 40 ng/µL rhC1rLP and start the reaction by adding 50 µL of Substrate Mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of Substrate Mixture.
- Read plate at 405 nm (absorbance) in kinetic mode for 5 minutes.
- 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) |
- rhC1rLP: 2 µg
- Z-GR-SBzl: 200 µM
- DTNB: 100 µM
Scientific Data
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Recombinant Human Complement Component C1rLP HA-tag His-tag (Catalog # 11614-CP) is 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).
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2 μg/lane of Recombinant Human Complement Component C1rLP HA-tag His-tag Protein (Catalog # 11614-CP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70‑76 kDa, under reducing conditions.
Background: Complement Component C1rLP
Recombinant human Complement C1r subcomponent-like protein (C1rLP), also known as C1r-like serine protease analog protein (CLSPa) is a 487 residue glycosylated, secreted serine peptidase-like protein (1, 2). It contains an N-terminal signal peptide, a CUB domain for substrate and protein-protein interaction, a truncated complement control protein module, and a trypsin-like serine protease domain with a characteristic catalytic triad but lacks an activation consensus peptide and an EGF domain found in homologous complement cascade proteases like C1r protein (1, 2). It is broadly expressed and secreted in tissues and abundantly found in liver, kidney, and myeloid cells where significant expression of serine proteases typically occurs (1). C1rLP has esterolytic activity on peptide thioesters with arginine at the P1 position but lower catalytic efficiency compared to C1r (2). It was reported to mediate cleavage of haptoglobin (3) and subsequently proposed to be of use in hemoglobin-binding therapeutics (4). Complement cascade serine proteases such as C1r play a role in innate immunity but the role of C1rLP in complement-mediated function is unclear as both activation and inhibition roles have been described in the literature (1, 2). C1rLP has been shown to be a pathological marker in immune microenvironments including within acute myocardial infarction and COVID19 as well as several types of cancer such as colon, leukemia, brain, endometrial, and melanoma (5-12). As a pathological marker, it has also been proposed as a potential therapeutic target for the treatment of these diseases (12).
- Lin, N. et al. (2004) Biochem. Biophys. Res. Commun. 321:329.
- Ligoudistianou, C. et al. (2005) Biochem. J. 387:165.
- Wicher, K.B. et al. (2004) Proc. Natl. Acad. Sci. 101:14390.
- Schaer, C.A. et al. (2018) BMC Biotechnol. 18:15.
- Liu, X. et al. (2021) ACS Omega 6:7951.
- Yang, S. et al. (2022) Front. Oncol. 12:934928.
- Chen J. et al. (2023) Aging 15:4444.
- Njoku K. et al. (2023) Br. J. Cancer 128:1723.
- Yoon, H.G. et al. (2023) PLoS One 18:e0295061.
- Beck. H.C. et al. (2024) Int. J. Mol. Sci. 25:2638.
- Liu, G. et al. (2024) Heliyon 11:e32337.
- Zhang, M. et al. (2024) Heliyon 10:e30616.
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