What is the theoretical molecular mass of Recombinant Human Coagulation Factor II/Thrombin Protein (catalog 1473-SE)?

The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the theoretical molecular mass of our product is ~35 kDa.

What is the C-terminal fragment mentioned in the Product Description?

The 14 kDa C-terminal fragment is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels and is the reason we report it.

The product page for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (Catalog # 1473-SE) lists three different molecular weights: light chain A, heavy chain B, and C-terminal fragment. What is the estimated molecular weight of this protein?

The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the molecular mass of our 1473-SE protein is ~ 35 kDa. The C-terminal fragment, 14 kDa, is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels which is the reason we report it.

Recombinant Human Coagulation Factor II/Thrombin Protein, CF

Name: Recombinant Human Coagulation Factor II/Thrombin Protein, CF
Brand: R&D Systems
SKU: 1473-SE
Rating: 5 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 1473-SE

Citations (3)

(1)

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Source:	NS0
Accession #:	P00734
Structure / Form:	Active form
Applications:	Enzyme Activity

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Product Specifications for Recombinant Human Coagulation Factor II/Thrombin Protein, CF

Source	Mouse myeloma cell line, NS0-derived human Coagulation Factor II/Thrombin protein Met1-Glu622 with a C-terminal 10-His tag The proenzyme was purified, activated and further purified.
Purity	>95%, 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	Thr328, Ile364 & Val515
Predicted Molecular Mass	4 kDa (light chain A), 31 kDa (heavy chain B), 14 kDa (C-terminal fragment)
SDS-PAGE	12 kDa, 21 kDa and 23 kDa, reducing conditions
Activity	Measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011). The specific activity is >20,000 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.

1473-SE

Formulation	Lyophilized from a 0.2 μm filtered solution in HEPES and NaCl.
Reconstitution	Reconstitute at 100 μg/mL in sterile 50 mM HEPES and 580 mM NaCl, pH 7.5.
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.

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Background: Coagulation Factor II/Thrombin

Coagulation Factor II, commonly known as thrombin, is an essential component of the coagulation cascade in which it converts fibrinogen to fibrin, activates factors V, VII, VIII, XIII and forms complexes with protein C and thrombomodulin (1). It also activates platelets and regulates the behavior of additional cells through protease‑activated receptors (PARs) (2). It may have either protective or deleterious functions, depending on the level and location (3). Its activity is regulated by endogenous inhibitors such as anti-thrombin III (serpin C1) or heparin cofactor II (serpin D1). A plasma serine protease, thrombin is synthesized in the liver as a 622 amino acid precursor with a 24 amino acid signal peptide. Cleavage by itself or by similar enzymes converts the proenzyme to three forms designated as alpha ‑, beta ‑ and gamma ‑thrombin. Composed of a disulfide bond-linked dimer of the light chain (A) (residues 328-363) and the heavy chain (B) (residues 364‑622), alpha -thrombin displays the diverse functions as described above. In comparison, the further processed B chains of beta - and gamma -thrombin have no known physiological function, but retain most of the activity towards small synthetic substrates (4). The recombinant human thrombin may be used to process proteins containing the thrombin cleavage site. The conditions for processing different proteins, such as the ratio of protein/thrombin, buffer components, temperature and time of incubation, may vary and should be empirically determined.

Item	Value
References	Degen, S.J. and E.W. Davie (1987) Biochemistry 26:6165. Coughlin, S.R. (2000) Nature 407:258. Xi, G. et al. (2003) J. Neurochem. 84:3. Rydel, T.J. et al. (1994) J. Biol. Chem. 269:22000.
Entrez Gene IDs	2147 (Human); 14061 (Mouse)
Alternate Names	Coagulation Factor II, Coagulation factor II receptor-like 2, Coagulation factor II receptor-like 2 (protease-actovated receptor 3), F2, PAR-3, PAR3proteinase-activated receptor 3, PT, Thrombin, Thrombin receptor-like 2, coagulation factor II (thrombin) receptor-like 2, protease-activated receptor 3, proteinase-activated receptor-3, serine protease

Citations for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (3)

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

Species: Mouse
Sample Types: Whole Cells
Applications: Cell Culture

KM Herum et al. (2020), Syndecan-4 Protects the Heart From the Profibrotic Effects of Thrombin-Cleaved Osteopontin J Am Heart Assoc, 9(3):e013518.
PMID: 32000579
Mogami et al. (2014), Effect of thrombin on human amnion mesenchymal cells, mouse fetal membranes, and preterm birth. J Biol Chem, 289(19):13295-307.
PMID: 24652285
Species: Human
Sample Types: Serum
Applications: ELISA Developmet

Wu FX et al. (2009), Identifying serological biomarkers of hepatocellular carcinoma using surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy. Cancer Lett., 279(2):163-70.
PMID: 19268439

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Recombinant Human Coagulation Factor II/Thrombin Protein, CF

Name: Anonymous

Application: Used thrombin for the growth of mamospheres.

Verified Customer | Posted 03/30/2019

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FAQs for Recombinant Human Coagulation Factor II/Thrombin Protein, CF

What is the theoretical molecular mass of Recombinant Human Coagulation Factor II/Thrombin Protein (catalog 1473-SE)?

The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the theoretical molecular mass of our product is ~35 kDa.
What is the C-terminal fragment mentioned in the Product Description?

The 14 kDa C-terminal fragment is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels and is the reason we report it.
The product page for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (Catalog # 1473-SE) lists three different molecular weights: light chain A, heavy chain B, and C-terminal fragment. What is the estimated molecular weight of this protein?

The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the molecular mass of our 1473-SE protein is ~ 35 kDa. The C-terminal fragment, 14 kDa, is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels which is the reason we report it.

Recombinant Human Coagulation Factor II/Thrombin Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

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Product Specifications for Recombinant Human Coagulation Factor II/Thrombin Protein, CF

Formulation, Preparation and Storage

Carrier Free

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Background: Coagulation Factor II/Thrombin

Citations for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (3)

Customer Reviews for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (1)

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Recombinant Human Coagulation Factor II/Thrombin Protein, CF

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