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H0393

Sigma-Aldrich

Hirudin

recombinant, expressed in unspecified host, ≥7,000 ATU/mg protein, powder

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About This Item

CAS Number:
8001-27-2
EC Number:
232-279-1
MDL number:
MFCD00145679
UNSPSC Code:
12352202
NACRES:
NA.77
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Product Name

Hirudin, recombinant, expressed in unspecified host, ≥7,000 ATU/mg protein (ATU = antithrombin units)

recombinant

expressed in unspecified host

Quality Level

200

form

powder

specific activity

≥7,000 ATU/mg protein (ATU = antithrombin units)

mol wt

~7 kDa

solubility

pyridine: soluble
water: soluble

shipped in

dry ice

storage temp.

−20°C

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Application

Antithrombotic efficacy by parenteral route is substantially prolonged by incorporation into cationic liposomes, which act as a sort of sustained-release device.

Physical properties

The anticoagulant hirudin is a ~7 kDa acidic protein containing 65 amino acid residues. It occurs naturally in leeches (Hirudo medicinalis). It is the most potent natural inhibitor of both soluble and clot-bound thrombin. Hirudin binds thrombin with high affinity and covers more than 20% of the surface area of thrombin, occluding both the active site and exosite I (fibrinogen and PAR recognition site). This coverage blocks thrombus growth and platelet activation. Hirudin is not metabolized in the bloodstream of humans and is eliminated unchanged via kidney filtration. Native hirudin contains a sulfated tyrosyl residue (Tyr63), three disulfide bridges, and a high proportion of dicarboxylic acids. Hirudin is not glycosylated, and lacks tryptophan, arginine, and methionine residues. At least 20 isoforms have been identified and sequenced.
This recombinant hirudin product corresponds to the HV-1 variant sequence, with the exception that the Tyr63 residue is not sulfated.

Unit Definition

One antithrombin unit (ATU) will neutralize one NIH unit of thrombin at 37 °C, based on direct comparison to an NIH thrombin reference standard.

Reconstitution

Hirudin is soluble in water. The literature cites the use of "Dilution Fluid II" (35.7 mM acetic acid, 35.7 mM sodium diethyl barbiturate, 0.85% NaCl, 1% bovine serum albumin, and 0.5% PEG, pH 7.4) to dissolve hirudin (500 ATU/mL) and thrombin. Hirudin is reported to be soluble in pyridine, but practically insoluble in alcohol, ether, acetone, or benzene.

Analysis Note

Protein determined by Lowry.

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Certificates of Analysis (COA)

Lot/Batch Number
0000322988
0000322988
SLCQ5268
SLCM2150
SLCK4656

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R Bischoff et al.
Biochemistry, 32(2), 725-734 (1993-01-19)
Natural hirudin variant 2 with a lysine residue in position 47 (rHV2-Lys47) was produced in a genetically engineered strain of Saccharomyces cerevisiae as a secreted protein of 65 amino acids and purified to greater than 99% homogeneity. Only reversed-phase high-performance
The complete amino acid sequence of hirudin, a thrombin specific inhibitor: Application of colour carboxymethylation.
Dodt, J., et al.
Febs Letters, 165, 180-184 (1984)
A Kudo et al.
Cerebrovascular diseases (Basel, Switzerland), 10(6), 424-430 (2000-11-09)
The role of thrombin as a spasmogen after subarachnoid hemorrhage was evaluated using the intrathecally administered thrombin inhibitor hirudin, released from a drug delivery system (DDS) based on collagen in a canine vasospasm model. The DDS was implanted into the
Serdar Duzgun et al.
Indian journal of plastic surgery : official publication of the Association of Plastic Surgeons of India, 47(1), 102-108 (2014-07-06)
The effect of recombinant hirudin, which is the most powerful antithrombotic agent, on flaps with venous insufficiency was investigated. Oedema and congestion are frequent on flaps, causing necrosis unpredictably. Venous insufficiency and thrombosis are experimentally and clinically more frequent than
Hirudin and hirudin-based peptides.
S R Stone et al.
Methods in enzymology, 223, 312-336 (1993-01-01)
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Questions

1–3 of 3 Questions  

  1. Hi Please help me for reconstituting of Hirudin H0393  H100UN . Can I solve it by distilled water ? How much of DW should be added to the vial? Another question is the vial seems to be empty however it is plumb . Is this because the amount of powder?

    1 answer

    1. This material is soluble in water, a rate has not been established. For assay purposes, this product is reconstituted at 10 units/100ul in 28.6 mM Barbital buffer (see product B5934) containing 1.0% (w/v) Bovine Serum Albumin and 0.5% (w/v) Polyethylene Glycol, pH 7.35. Please see the link below to review the product datasheet which includes detailed preparation instructions.
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/320/022/h0393pis.pdf

      The activity specification for this product is minimum 7000 units per milligram protein. Occasionally, a small amount of powder may not be visible. A gentle spin or ‘shake down’ of the vial may move the powdered product to the bottom of the vial.

      Helpful?

  2. Thanks for the previous answer, but the "units" definition is confusing. Does "100-units pack" mean 100-IU (in the International Standard system) or -ATU again?

    1 answer

    1. One antithrombin unit (ATU) will neutralize one NIH unit of thrombin at 37°C, based on a direct comparison to an NIH thrombin reference standard. There is no International Standard Unit value associated with this product.

      Helpful?

  3. Hi, some years ago we bought H0393-100UN but lost the info sheet. How many mg protein does the single vial contain? Thank you very much, Dario

    1 answer

    1. A link to the general product information sheet is provided on this product page. See the link below to review. You may either print or save a copy as needed.
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/320/022/h0393pis.pdf

      The specific activity will determine the amount of protein in each vial. This value will be listed on the lot-specific Certificate of Analysis. Please see the link below to access the Certificate of Analysis for the currently available lot: https://www.sigmaaldrich.com/certificates/COFA/H0/H0393/H0393-100UN_______SLCK4656__.pdf

      As the specific unit activity is batch dependent, the currently available lot, SLCK4656, has a units/mg activity of 9769. Thus, the 100-unit pack size for this lot contains 0.01024 mg protein.

      Please refer to the product label to verify your lot number. Should a different lot number be needed please navigate to the link https://www.sigmaaldrich.com/techservice, click on "Product Technical Inquiries" under the Products Section with all the required information so that a member of the Technical Service team can reach out to assist further. Thank you.

      Helpful?

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