Recombinant Mouse Coagulation Factor XI Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

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.
Source
Mouse myeloma cell line, NS0-derived mouse Coagulation Factor XI protein
Glu19-Val624, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Glu19
Structure / Form
Disulfide-linked homodimer
Protein/Peptide Type
Recombinant Enzymes
Gene
F11
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
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.
SDS-PAGE
74 kDa, reducing conditions
Publications
Read Publication using
4556-SE in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • 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
  1. Dilute rmFactor XI to 100 µg/mL in Activation Buffer containing 10 µg/mL of Thermolysin.
  2. Incubate at 37 °C for 1 hour.
  3. Stop reaction with EDTA at a final concentration of 40 mM.
  4. Dilute incubated rmFactor XI to 2 ng/µL in Assay Buffer.
  5. Dilute Substrate to 200 µM in Assay Buffer.
  6. 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.
  7. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  8. 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).

Per Well:
  • rmFactor XI:  0.100 µg
  • Thermolysin: 0.010 µg
  • Substrate: 100 µM

Notes

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
  • EC 3.4.21
  • EC 3.4.21.27
  • FXIPlasma thromboplastin antecedent
  • MGC141891
  • PTA

Background

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.

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

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Publications for Coagulation Factor XI (4556-SE)(1)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 1 application: DirELISA.


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Bioinformatics

Gene Symbol F11
Uniprot