Recombinant Human Coagulation Factor Xa Protein, CF

Name: Recombinant Human Coagulation Factor Xa Protein, CF
Brand: rnd
SKU: 1063-SE
Rating: 5 (1 reviews)

Catalog # 1063-SE

Citations (2)

(1)

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Key Product Details

Source:	Sf 21 (baculovirus)
Accession #:	P00742
Structure / Form:	Active form
Applications:	Enzyme Activity

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Product Specifications

Source	Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Coagulation Factor X protein Leu24-Lys488 with a C-terminal 10-His tag Proform Factor X was expressed, purified, activated and further purified to yield Factor Xa.
Purity	>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin level	<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Tyr84, Phe124 & Ile235
Predicted Molecular Mass	12 kDa & 30 kDa
SDS-PAGE	13-14 kDa and 33-36 kDa, reducing conditions
Activity	Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH₂ (Catalog # ES002). The specific activity is >700 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation and Storage

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.

1063-SE

Formulation	Lyophilized from a 0.2 μm filtered solution in MES, NaCl and CaCl₂.
Reconstitution	Reconstitute at 100 μg/mL in sterile 25 mM MES, 150 mM NaCl and 5 mM CaCl₂, pH 6.0.
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 reconstitution.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Assay Procedure

Materials

Assay Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% Brij-35, pH 7.5 (TCNB)
Recombinant Human
Coagulation Factor Xa (rhFactor Xa) (Catalog # 1063-SE)
Substrate: Fluorogenic Peptide Substrate II, MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH₂ (Catalog # ES002)
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

Dilute rhFactor Xa to 0.4 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
Load 50 µL of the 0.4 ng/µL rhFactor Xa into plate and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rhFactor Xa.
Read at excitation and emission wavelengths of 320 nm and 405 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 using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975)

Per Well:

rhFactor Xa: 0.02 µg
Substrate: 10 µM

Background: Coagulation Factor X

As the only known physiological activator of thrombin, Factor X is a vitamin K-dependent plasma protease that plays a pivotal role in blood coagulation. Human Factor X is initially synthesized in the liver as a single-chain precursor of 488 amino acid residues with a signal peptide and a pro region (residues 1‑40). Both the intrinsic and extrinsic pathways activate Factor X to Xa, which consists of light (residues 41‑179) and heavy (residues 235‑488) chains linked by a disulfide bond. The light chain contains a Gla and two EGF-like domains and the heavy chain corresponds to the serine protease domain. The full-length human Factor X was expressed and the pro enzyme was purified and activated. The active protease (rhXa) was further purified and analyzed for its activity towards either peptides or proteins containing a Xa cleavage site. In addition to the activity described in the Activity Assay Protocol, rhFX also be used to cleave fusion proteins containing a Factor Xa cleavage site. The conditions for the optimal cleavage of a particular fusion protein, such as the molar ratio between rhFX and the protein and time and temperature of incubation, are protein-dependent and need to be individually determined.

Item	Value
Entrez Gene IDs	2159 (Human); 14058 (Mouse); 29243 (Rat)
Alternate Names	Cf10, Coagulation Factor X, EC 3.4.21, EC 3.4.21.6, F10, FX, FXa, Prothrombinase, Stuart Factor, Stuart-Prower factor, factor Xa

Citations (2)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human Coagulation Factor Xa Protein, CF include:

Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay

Amara U et al. (2010), Molecular intercommunication between the complement and coagulation systems. J. Immunol., 185(9):5628-36.
PMID: 20870944
Species: Human
Sample Types: Recombinant Protein
Applications: Fc Cleavage

Zimmer G et al. (2008), Ephrin-A5 acts as a repulsive cue for migrating cortical interneurons. Eur. J. Neurosci., 28(1):62-73.
PMID: 18662335

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Recombinant Human Coagulation Factor Xa Protein, CF

Name: Anonymous

Application: Enzymatic activity in vitro

Verified Customer | Posted 08/03/2016

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Recombinant Human Coagulation Factor Xa Protein, CF

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Key Product Details

Complete Your Experiment

Complete Your Research

Product Specifications

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Assay Procedure

Materials

Background: Coagulation Factor X

Citations (2)

Customer Reviews (1)

Recombinant Human Coagulation Factor Xa Protein, CF

Product Datasheets

Product Datasheets

COA

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SDS

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