Recombinant Human Furin His-tag Protein, CF Summary
Details of Functionality |
Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (Catalog # ES013). The specific activity is >125 pmol/min/μg, as measured under the described conditions. |
Source |
Human embryonic kidney cell, HEK293-derived human Furin protein Asp108-Ala574, with a C-terminal 6-His tag |
Accession # |
|
N-terminal Sequence |
Asp108 |
Structure / Form |
Mature |
Protein/Peptide Type |
Recombinant Enzymes |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note |
<0.10 EU per 1 μg of the protein by the LAL method. |
Applications/Dilutions
Dilutions |
|
Theoretical MW |
52 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
SDS-PAGE |
44-66 kDa, under reducing conditions |
Packaging, Storage & Formulations
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.
|
Buffer |
Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl, Brij-35 and Glycerol. |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Assay Procedure |
- Assay Buffer: 25 mM Tris, 1 mM CaCl2, 0.5% (w/v) Brij-35, pH 9.0
- Recombinant Human Furin (rhFurin) (Catalog # 11338-SE)
- Substrate: p-Glu-Arg-Thr-Lys-Arg-AMC
(Catalog #
ES013), 8 mM stock in DMSO
- Black 96-well Plate
- Plate Reader with Fluorescence Read Capability
- Dilute rhFurin to 4 µg/mL in Assay Buffer.
- Dilute Substrate to 100 µM in Assay Buffer.
- Load into a black 96-well plate 50 µL of 4 µg/mL rhFurin, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 100 µM Substrate.
- Read plate at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Calculate specific activity:
Specific Activity (pmol/min/µg) = | Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) | amount of enzyme (µg) |
*Adjusted for Substrate Blank **Derived from calibration standard 7-amino, 4-Methyl Coumarin
Per Well: - rhFurin: 0.2 µg
- Substrate: 50 µM
|
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Furin His-tag Protein, CF
Background
Furin is a member of the proprotein convertase (PC) family, which belongs to the subtilisin superfamily of serine protease (1-3). As a cellular protease, Furin cleaves after Arg-Xaa-Lys/Arg-Arg-like motifs typically at the end of the pro region in a variety of proproteins within the secretory pathway including growth factors and receptors, extracellular matrix proteins, and other proteases. Through regulation of proprotein maturation, Furin has an essential role in embryogenesis and homeostasis and is implicated in various pathologies such as cancer (4), neurodegenerative diseases (5) and infectious disease (6). Furin is synthesized as a 794 amino acid type I transmembrane protein precursor with a signal peptide (residues 1-24), a pro region (residues 25-107) that plays a crucial role in the folding, activation and transport of Furin, and a mature chain (residues 108-794) (1-3). The mature chain consists of the subtilisin-like catalytic domain, a P domain, which is essential for enzyme activity and the modulation of pH and calcium requirements, and a cytoplasmic domain, which controls the localization and sorting of Furin in the trans-Golgi network/endosomal system. The purified recombinant human Furin corresponds to a truncated form of the mature enzyme terminated before the transmembrane domain.
- Van den Ouweland, A.M. et al. (1990) Nucleic Acids Res. 18:664.
- Barr, P.J. et al. (1991) DNA Cell Biol. 10:319.
- Thomas, G. (2002) Nature Rev. Mol. Cell Biol. 3:753.
- He, Z. et al. (2022) Oncogene. 9:1252.
- Zhang, Y. et al. (2022) Transl. Neurodegener. 11:39.
- Braun, E. and D. Sauter (2019) Clin. Transl. Immunology. 8:e1073.
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