| Reactivity | All-MultiSpecies Glossary |
| Applications | ELISA |
| Standard Curve Range | 1 - 300 ng/ml (0.1 - 30 ng/well) |
| Sensitivity | 3 ng/ml |
| Description | This assay works on the sandwich-ELISA principle and uses the K1 (IgG2a) mouse monoclonal antibody to dsRNA as a catcher antibody. The monoclonal antibody K2 (IgM) is used as the detector antibody. Based on the use of two double-stranded RNA (dsRNA)-specific monoclonal antibodies the dsRNA Detection Kit allows sensitive and selective detection of dsRNA molecules (larger than 30-40 bp), independent of their nucleotide composition and sequence. The detection is highly specific: dsRNA can be detected in nucleic acid extracts in the presence of 1,000-10,000-fold excess of other nucleic acids. Source The two dsRNA antibodies used in this kit were produced as follows: 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 clones K1 and K2. |
| Specificity | We recommend using the kit to detect viral dsRNAs or large natural or synthetic dsRNAs of non-viral origin in nucleic acid extracts, as well as to detect the presence of undesired dsRNA molecules in artificially synthesized (m)RNA preparations. Serial dilutions of the Poly(I:C) dsRNA standard (included in the kit) can be used as a positive control. For the exact detection protocol we refer to the kit manual that can be downloaded from our website Caution: The Poly(I:C) dsRNA positive control included in this sandwich ELISA kit is not intended to be used as a quantitative standard for other dsRNA preparations. Low molecular weight polyinosine-polycytidylic acid (i.e. poly(I:C)) is a synthetic analog of double-stranded RNA (dsRNA) and is comprised of short strands of inosine poly(I) homopolymer annealed to strands of cytidine poly(C) homopolymer. The average size of our Poly(I:C) LMW is 0.2 kb to 1 kb. The anti dsRNA antibodies J2 and/or K2 used in this kit may exhibit a different degree of reactivity with different dsRNAs obtained from synthetic or natural sources. It is therefore only intended to be used as a positive control to see if the ELISA has been executed correctly and that the test shows a linear relationship between the amount of dsRNA and the read out for the OD450. |
| Standard Curve Range | 1 - 300 ng/ml (0.1 - 30 ng/well) |
| Sensitivity | 3 ng/ml |
| Assay Type | Sandwich ELISA |
| Inter-Assay | 5 - 10% |
| Intra-Assay | 5 - 10% |
| Sample Volume | 100 ul |
| Kit Type | ELISA Kit (Colorimetric) |
| Storage | Store at 4C short term. Store at -20C long term. Avoid freeze-thaw cycles. |
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