Anti Hirudin
Mouse monoclonal antibody
Cat.No. HYB 010-02
Preparation: Protein-A/G purified
Content: Available in 200 µL and 1 mL
Concentration: 1 mg/mL
Solvent: 0.01 M phosphate buffer, pH 7.4, with 0.5 M NaCl and 15mM sodium azide
Storage: In the dark at 4-8ºC
Hirudin is a highly effective and very specific anticoagulant which specifically inhibits formation of fibrin and activation of factors V, VIII and XIII by forming irreversible, noncovalent 1:1 complexes with a-thrombin. Hirudin is a 7 kDa protein naturally originating from the medical leech Hirudo medicinalis.
Recombinant hirudin (rHV2) produced in S. cerevisiae and coupled to diphteria toxoid.
The affinity (Kd) of HYB 010-02 for rHV2K47 is 3.5x10E-8 M (1). Crossreactivity to hirudin added to rabbit, rat, dog and monkey plasma.
The epitope of HYB 010-02 is located at the C-terminal part of hirudin. It differs from that of HYB 010-05 and HYB 010-06 as determined by sandwich ELISA.
In Western blotting after SDS-PAGE, HYB 010-02 reacts with full length and C-terminal derivatized hirudins in both reduced and unreduced forms. HYB 010-02 is suitable as capture antibody in sandwich ELISA with HYB 010-06 as detection antibody on hirudin from plasma or urine (1). Binding of HYB 010-02 to hirudin inhibits the interaction with a-thrombin.
| Method | Usability | Dilution guideline | References |
| ELISA | Yes | 1:4000 | 1 |
| Immunoblotting | Yes | 1 | |
| Immunohistochemistry | Not determined |
1. Koch C, Whitechurch O, Cordier P, Roitsch C (1993) Anti-hirudin monoclonal antibodies directed toward discontinuous epitopes of the hirudin amino-terminal and epitopes involving the carboxy-terminal hirudin amino acids. Anal Biochem 214:301-312.
2. Skrzypczak-Jankun E, Carperos VE, Ravichandran KG, Tulinsky A, Westbrook M, Maraganore JM (1991) Structure of the hirugen and hirulog 1 complexes of alpha-thrombin. J Mol Biol 221:1379-1393.
3. Rydel TJ, Tulinsky A, Bode W, Huber R (1991) Refined structure of the hirudin-thrombin complex. J Mol Biol 221:583-601.
