Recombinant Human Furin His-tag Protein, CF

Catalog #: 11338-SE Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
11338-SE-025

Recombinant Human Furin His-tag Protein Enzyme Activity.

1 Image

Product Details

Supplemental Products

Reviews

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Furin His-tag Protein, CF Summary

Product Specifications

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.

Activity

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 #

P09958.2

N-terminal Sequence
Analysis

Asp108

Structure / Form

Mature

Predicted Molecular Mass

52 kDa

SDS-PAGE

44-66 kDa, under reducing conditions

Product Datasheets

11338-SE

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

11338-SE

Formulation	Supplied as a 0.2 μm filtered solution in Tris, CaCl₂, NaCl, Brij-35 and Glycerol.
Shipping	The product is shipped with dry ice or equivalent. 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.

Assay Procedure

Materials

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 V_max* (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

Scientific Data

Enzyme Activity

View Larger

Recombinant Human Furin (Catalog # 11338-SE) is measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (ES013).

Reconstitution Calculator

Background: Furin

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.

References

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.

Entrez Gene IDs

5045 (Human); 18550 (Mouse)

Alternate Names

Dibasic-processing enzyme; EC 3.4.21; EC 3.4.21.75; FUR; FURdibasic processing enzyme; furin (paired basic amino acid cleaving enzyme); Furin; furin, membrane associated receptor protein; PACE; PACEFES upstream region; paired basic amino acid cleaving enzyme (furin, membrane associated receptorprotein); Paired basic amino acid residue-cleaving enzyme; PCSK3; PCSK3furin; proprotein convertase subtilisin/kexin type 3; SPC1