Mouse anti double-stranded RNA (K2)

Over the past decade our double-stranded RNA (dsRNA)antibodies have been used extensively to detect and characterise plant and animal viruses with dsRNA genomes or intermediates. In addition, the anti-dsRNA antibodies can be used as a diagnostic tool to detect pathogens, including detection in paraffin-embedded fixed tissue samples (Richardson et al. 2010). _x000D_ K2 anti-dsRNA monoclonal antibody has an IgM isotype._x000D_ _x000D_ K2 has primarily been used in ELISA and sandwich ELISA. Generally, K2 can be used in applications where an anti-dsRNA antibody with an isotype other than IgG (IgG2a) is required.

Female DBA/2 mice were injected intraperitonially with a mixture of 50 ug L-dsRNA and 75 ug methylated bovine serum albumin, emulsified in complete Freund's adjuvant. After several boosts spleen cells were fused with Sp2/0-Agl4 myeloma cells to generate the hybridoma clone.

MAb K2 is primarily used for a sandwich ELISA to detect and quantitate (after calibration) dsRNA (see Schönborn et al.). For this application it should be diluted with PBS. It may also be advantageous to use K2 for immunofluorescence studies. The optimum working dilution of the antibody for any specific application should be established by titration._x000D_ _x000D_ Not for use for clinical purposes. For in vitro use only.

The mAb K2 recognises double-stranded RNA (dsRNA) provided that the length of the helix is greater than or equal to 40 bp dsRNA; Recognition is independent of the sequence and nucleotide composition of the antigen. All naturally occurring dsRNAs investigated up to; Now (4050 species) as well as poly(I).poly(C) and poly(A).poly(U) have been recognised by K2. As described by Schönborn et al. K2 binds with high avidity to all dsRNAs investigated. // Culture supernatant (RPMI, 5% fetal calf serum)

1) Schönborn, J., Oberstrass, J., Breyel, E., Tittgen, J., Schumacher, J. and Lukacs, N. (1991) Monoclonal antibodies to double-stranded RNA as probes of RNA structure in crude nucleic acid extracts. Nucleic Acids Res.19, 2993-3000. _x000D_ _x000D_ 2) Lukacs, N. (1994) Detection of virus infection in plants and differentiation between coexisting viruses by monoclonal antibodies to double-stranded RNA. J. Virol. Methods 47, 255-272. _x000D_ _x000D_ 3) Lukacs, N. (1997) Detection of sense:antisense duplexes by structurespecific anti-RNA antibodies. In: Antisense Technology. A Practical Approach, C. Lichtenstein and W. Nellen (eds), pp. 281-295. IRL Press, Oxford._x000D_ _x000D_ 4) S. J. Richardson, A. Willcox, D. A. Hilton, S. Tauriainen, H. Hyoty, A. J. Bone, A. K. Foulis, N. G. Morgan. Use of antisera directed against dsRNA to detect viral infections in formalin-fixed paraffin-embedded tissue. J Clin Virol. (2010) 49(3); 180-5. doi: 10.1016/j.jcv.2010.07.015.

After delivery antibodies should be aliquoted and stored at -20 ° or -70 °C; After adding 10 mM sodium azide undiluted antibody can also be stored at +4 °C for a short period of time; For long term storage the mAb should be kept frozen; Repeated freezing/thawing cycles should be avoided;