Furin/PCSK3 His Tag Protein, Human
SKU: 76462305441

Furin/PCSK3 His Tag Protein, Human

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Description

Furin/PCSK3 His Tag Protein, HumanProduct Specification Species Human Synonyms FUR,PCSK3,SPC1,PACE Accession P09958 1 Amino Acid Sequence Gln27 Ala574 with His Tag at the C Terminus Expression System HEK293 Molecular Weight 55 72kDa (Reducing) Purity 90% by SDS PAGE Conjugation Unconjugated Tag His Tag Physical Appearance Lyophilized powder Storage Buffer PBS, PH7. 4, 5% trehalose Reconstitution Reconstitute at 0. 1 1 mg ml according to the size in ultrapure water after rapid

Product Specification


Species Human
Synonyms FUR,PCSK3,SPC1,PACE
Accession P09958-1
Amino Acid Sequence

Gln27-Ala574 with His Tag at the C-Terminus

Expression System HEK293
Molecular Weight

55-72kDa (Reducing)

Purity >90% by SDS-PAGE
Conjugation Unconjugated
Tag His Tag
Physical Appearance Lyophilized powder
Storage Buffer

PBS, PH7.4, 5% trehalose

Reconstitution

Reconstitute at 0.1-1 mg/ml according to the size in ultrapure water after rapid centrifugation.

Stability & Storage

· 12 months from date of receipt, lyophilized powder stored at -20 to -80℃.
· 3 months, -20 to -80℃ under sterile conditions after reconstitution.
· Please do not repeated freeze-thaw cycles.

Reference

1.Anderson ED, Molloy SS, Jean F, Fei H, Shimamura S, Thomas G. The ordered and compartment-specfific autoproteolytic removal of the furin intramolecular chaperone is required for enzyme activation. J Biol Chem. 2002 Apr 12;277(15):12879-90.
2.Dahms SO, Hardes K, Becker GL, Steinmetzer T, Brandstetter H, Than ME. X-ray structures of human furin in complex with competitive inhibitors. ACS Chem Biol. 2014 May 16;9(5):1113-8.

Background

Furin, also known as Paired Basic Amino Acid Cleaving Enzyme (PACE), is a key member of the subtilisin-like proprotein convertase family. Functioning as a calcium-dependent serine protease, its structure includes an N-terminal signal peptide, a prodomain, a catalytic domain, a P-domain responsible for maintaining enzymatic activity and calcium/pH sensitivity, a transmembrane region, and a cytoplasmic tail. Primarily localized within the Golgi apparatus/endosomal system, it activates a wide range of proprotein substrates—such as growth factors, receptors, extracellular matrix proteins, and pathogen proteins (e.g., envelope proteins of HIV and influenza virus)—by specifically cleaving after the Arg-X-(Arg/Lys)-Arg sequence motif. It plays a critical role in embryogenesis, peripheral immune tolerance, and cellular homeostasis. Dysregulation of its function is closely linked to tumor progression, neurodegenerative diseases, and various pathogen infections, making it a significant therapeutic target in clinical applications.

Protocol

Assay protocol
Principle: Measured by its ability to cleave the fluorogenic peptide substrate, Pyr-Arg-Thr-Lys-Arg-AMC.
Materials
Assay buffer: 25 mM Tris, 1 mM CaCl2, pH 9.0
Furin/PCSK3 His Tag Protein, Human (UA011272)
Substrate: Pyr-Arg-Thr-Lys-Arg-AMC (abs45133051)
96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)
Plate Reader (PerkinElmer, excitation 380 nm and emission 460 nm)
Produce
Dilute Human Furin Protein to 2 µg/mL/4 µg/mL/8 µg/mL in Assay Buffer.
Dilute Substrate to 100 µM in Assay Buffer.
Load into a black well plate 50 µL of 2 µg/mL/4 µg/mL/8 µg/mL of Human Furin Protein, 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 at excitation and emission wavelengths of 380 nm and 460 nm, respectively in kinetic mode for 10 minutes.
Calculate specific activity:
*Adjusted for Substrate Blank
**Derived using calibration standard 7-amino-4-Methyl Coumarin
Conversion Factor (pmol/RFU)
Produce
Buffer: Assay buffer
Calibration standard: standard 7-amino-4-Methyl Coumarin (Enzo, Catalog # BML-KI107-0001).
Add 100 µL Dilute Calibration standard, Standard curve 1500/750/375/188/94/47/24/12/6/3 pmol per well or blank (Assay buffer) per well.
Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively in endpoint mode.
Linear Regression of 7-amino-4-Methyl Coumarin (pmol) (y)—RFU-Blank(x).
Note
Storage Conditions: After reconstitution, aliquot and store at -80°C. Please do not
repeated freeze-thaw cycles.

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SKU: 76462305441

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