Recombinant Mouse Coagulation Factor XI Protein, CF

Catalog #: 4556-SE
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
4556-SE-010

R&D Systems Recombinant Proteins and Enzymes

1 Image

All Coagulation Factor XI Proteins and Enzymes

Product Details

Citations (1)

Supplemental Products

Reviews

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

Recombinant Mouse Coagulation Factor XI Protein, CF Summary

Product Specifications

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.

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, t-butyloxycarbonyl-Ile-Glu-Gly-Arg-7-amido-4-methylcoumarin (Boc-IEGR-AMC). The specific activity is >100 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Coagulation Factor XI protein
Glu19-Val624, with a C-terminal 10-His tag

Accession #

NP_082342

N-terminal Sequence
Analysis

Glu19

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

69 kDa

SDS-PAGE

74 kDa, reducing conditions

Product Datasheets

4556-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.

4556-SE

Formulation	Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
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, -70 °C as supplied. 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials

Activation Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
Assay Buffer: 50 mM Tris, 1 mM EDTA, pH 7.5
Recombinant Mouse Coagulation Factor XI (rmFactor XI) (Catalog # 4556-SE)
Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
EDTA, pH 8.0 (Sigma, Catalog # E-4884), 0.5 M stock in deionized water
Substrate: BOC-Ile-Glu-Gly-Arg-AMC (Bachem, Catalog # I-1100), 10 mM stock in DMSO
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

Dilute rmFactor XI to 100 µg/mL in Activation Buffer containing 10 µg/mL of Thermolysin.
Incubate at 37 °C for 1 hour.
Stop reaction with EDTA at a final concentration of 40 mM.
Dilute incubated rmFactor XI to 2 ng/µL in Assay Buffer.
Dilute Substrate to 200 µM in Assay Buffer.
Load 50 µL of 2 ng/µL rmFactor XI into a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate.
Read 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 using calibration standard 7-amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).

Per Well:

rmFactor XI: 0.100 µg
Thermolysin: 0.010 µg
Substrate: 100 µM

Reconstitution Calculator

Background: Coagulation Factor XI

Coagulation factors XI and XIa refer to the pro and active forms of the same protease, respectively (1). Factor XI is synthesized in the liver and circulates in the plasma as a disulfide bond-linked dimer complexed with high molecular weight kininogen. Factor XI is converted into XIa via either the contact phase of blood coagulation or thrombin-mediated activation on the platelet surface. The resulting XIa converts factor IX into IXa, which subsequently activates factor X into Xa. Factor Xa in turn activates factor II/thrombin to complete the coagulation cascade. Patients with factor XI deficiency are prone to excessive bleeding after hemostatic challenge. There are two alternative splicing forms. Isoform 1 corresponds to the circulating plasma factor XI and isoform 2 is produced by platelets and megakaryocytes but absent from other blood cells (2). The 624 amino acid precursor of isoform 1 consists of a signal peptide (residues 1 to 18) and the mature chain (residues 19 to 624). The mature chain (XI) can be further processed into the heavy chain (residues 19 to 389) and the light chain (residues 390 to 624) (XIa). The purified rmFactor XI corresponds to isoform 1 (residues 19 to 624), which can be activated by treatment with thermolysin under the conditions described in the Activity Assay Protocol.

References

Wash, P.N. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. p. 1651.
Hsu, T.C. et al. (1998) J. Biol. Chem. 273:13787.

Entrez Gene IDs

2160 (Human); 109821 (Mouse)

Alternate Names

Coagulation Factor XI; EC 3.4.21; EC 3.4.21.27; FXIPlasma thromboplastin antecedent; MGC141891; PTA

Related Research Areas

Citation for Recombinant Mouse Coagulation Factor XI Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

Coagulation Factor XI vaccination: an alternative strategy to prevent thrombosis
Authors: Rongxiu Li
J. Thromb. Haemost., 2016-12-24;0(0):.
Species: Mouse
Sample Types: Serum
Applications: Direct ELISA