Recombinant Human Complement Component C1rLP His Protein, CF

R&D Systems | Catalog # 11614-CP

HA-tag His-tag
R&D Systems
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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Complement Component C1rLP His Protein (11614-CP)
  • 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

CHO

Accession Number

NP_057630.1

Applications

Binding Activity
View all Complement Component C1rLP Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Complement Component C1rLP protein
Cys36-Asp487 with N-terminal HA (YPYDVPDYA) and 6-His tags

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr

Predicted Molecular Mass

52 kDa

SDS-PAGE

70-76 kDa, under 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 >30 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human Complement Component C1rLP His Protein, CF

Recombinant Human Complement Component C1rLP HA-tag His-tag Protein Enzyme Activity.

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).

Recombinant Human Complement Component C1rLP HA-tag His-tag Protein SDS-PAGE.

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.

Formulation, Preparation, and Storage

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.
  • 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 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).

References

  1. Lin, N. et al. (2004) Biochem. Biophys. Res. Commun. 321:329.
  2. Ligoudistianou, C. et al. (2005) Biochem. J. 387:165.
  3. Wicher, K.B. et al. (2004) Proc. Natl. Acad. Sci. 101:14390.
  4. Schaer, C.A. et al. (2018) BMC Biotechnol. 18:15. 
  5. Liu, X. et al. (2021) ACS Omega 6:7951.
  6. Yang, S. et al. (2022) Front. Oncol. 12:934928.
  7. Chen J. et al. (2023) Aging 15:4444.
  8. Njoku K. et al. (2023) Br. J. Cancer 128:1723.
  9. Yoon, H.G. et al. (2023) PLoS One 18:e0295061.
  10. Beck. H.C. et al. (2024) Int. J. Mol. Sci. 25:2638.
  11. Liu, G. et al. (2024) Heliyon 11:e32337.
  12. Zhang, M. et al. (2024) Heliyon 10:e30616.

Long Name

Complement C1r Subcomponent-like Protein

Alternate Names

C1RL, C1RL1, CLSPa

Entrez Gene IDs

51279 (Human); 232371 (Mouse)

Gene Symbol

C1RL

UniProt

NP_057630.1

Additional Complement Component C1rLP Products

Product Documents for Recombinant Human Complement Component C1rLP His Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Complement Component C1rLP His Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Complement Component C1rLP His Protein, CF (11614-CP):

Materials
  • 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
  1. Dilute rhC1rLP to 40 ng/µL in Assay Buffer.
  2. Prepare a Substrate Mixture containing 400 µM of Z-GR-SBzl and 200 µM of DTNB in Assay Buffer.
  3. 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. 
  4. Read plate at 405 nm (absorbance) 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 of 0.320 cm
Per Well:
  • rhC1rLP: 2 µg
  • Z-GR-SBzl: 200 µM 
  • DTNB: 100 µM





















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