Recombinant Mouse Coagulation Factor XI Protein, CF Summary
Details of Functionality
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.
Mouse myeloma cell line, NS0-derived mouse Coagulation Factor XI protein Glu19-Val624, with a C-terminal 10-His tag
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
<1.0 EU per 1 μg of the protein by the LAL method.
69 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.
74 kDa, reducing conditions
Read Publication using 4556-SE in the following applications:
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 Vmax* (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).
rmFactor XI: 0.100 µg
Thermolysin: 0.010 µg
Substrate: 100 µM
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Mouse Coagulation Factor XI Protein, CF
Coagulation Factor XI
FXIPlasma thromboplastin antecedent
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.
Wash, P.N. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. p. 1651.
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