murine MAb against human factor VIII, light chain
Description
The monoclonal antibody (clone ESH-4) is directed against human factor VIII:C. Mice were immunized with purified human factor VIII:C cryoprecipitate. Hybridomas were created by fusion of the immunized spleen cells with NS-1 murine myeloma cell lines.
Properties
The antibody (subclass IgG2a) is directed against the light chain, aa: 2303-2332, phospholipid binding site on C-terminus.
The cross-reactivity was tested via IRMA, in comparison with human plasma (100%): the antibody does not cross react with Pig, Dog, Guinea Pig, and Cat, the antibody does cross-react with Rabbit (145%) and Baboon (112%).
Presentation
Screw capped vial containing 0.5 mg of purified antibody in PBS, pH 7.2, 0.1 % ProClin. The IgG concentration is 1 mg/ml. Spin the vial briefly before opening.
Storage and Stability
Store the antibody at 2°-8°C. For long-term storage the antibody should be aliquoted and stored at –20°C or colder. It is recommended to avoid freeze-thaw cycles.
Applications
A. Immunopurification / Immunodepletion
The antibody binds factor VIII:C from plasma when covalently coupled to gels (1).
B. Immunofluorescence Microscopy
The antibody is useful for immunofluorescence (3).
C. Inhibitor
Clot inhibition was tested by incubation for 2 hrs
at 37°C. 89% inhibition at 3 µg/mL (highest conc. tested). Recombinant human factor VIII binding
to vWF is inhibited at about 3 µg/mL (Per Dr. E. Saenko, ARC, Rockville, MD).
The antibody inhibits FVIIIa binding to platelets (5,6), platelet-dependent factor Xase activity is inhibited 70% to 85% by the antibody (6).
The antibody is conformation dependent and is not useful for Western blotting.
Category: Research use only
Type: Antibody
Product Availability: Worldwide
Manufacturer: ImmBioMed GmbH & Co KG, Germany
For more information please click .pdf icon below.
murine MAb against human factor VIII, light chain, clone ESH-4
Cat.No. ADGESH-4
Article no.: 938964
Unit: 0,5 mg
Code: ADGESH-4
Manufacturer: ImmBioMed GmbH & Co. KG
References
- The production and characterisation of a panel of ten murine monoclonal antibodies to human procoagulant factor VIII. Griffin, B. D., et al. Thromb Haemost, 1986; 55: 40-46.
- Development, optimization and use of an enzyme linked immunosorbent assay (ELISA) to measure factor VIII antigen utilizing monoclonal antibodies. Hornsey, V. S., et al. Transf Med, 1992; 2: 223-229.
- In vivo production of human factor VII in mice after intrasplenic implantation of primary fibroblasts transfected by receptor-mediated, adenovirus-augmented gene delivery. Zatloukal, K., et al. Proc Nat Acad Science USA, 1994; 91: 5148-5152.
- Some factor VIII inhibitor antibodies recognize a common epitope corresponding to C2 domain amino acids 2248 through 2312, which overlap a phospholipid-binding site. Scandella, D., et al. Blood, 1995; 86: 1811-1819.
- Role of the C2 domain of factor VIIIa in the assembly of factor-X activating complex on the platelet membrane. Ahmad SS, and Walsh PN. Biochem, 2005; 44(42):13858-13865.
- Platelet binding sites for factor VIII in relation to fibrin and phosphatidylserine. Gilbert GE et al., Blood. 2015; 126(10):1237-1244.