Immunofluorescence

All three of our antibodies (J2, K1 and K2) have been used for the immunhistological detection of dsRNA.

This is the primary use of these antibodies, documented by almost 200 publications. Histological preparations (suitably fixed cultured cells or thin sections of tissue) are incubated with one of our antibodies. The antibodies bind any dsRNA present with high affinity, and the bound anti-dsRNA antibody is then visualised using a labelled secondary antibody (indirect immunostaining). The secondary antibody is specific for mouse IgG2a (for J2 or K1) or IgM (for K2) and derivatised with a fluorescent dye or an enzyme.

Immunofluorescence microscopy using J2 antibody reveals dsRNA (labelled in red), a marker for the the viral replication complex, in HuH-7 cells infected with Dengue virus. Cellular DNA is labelled with DAPI (blue). Figure taken from Anwar et al. (2011) PLoS One 6:e23246.


Indirect immunofluorescence has been carried out using a variety of fluorescent labels (FITC, Rhodamine, various Alexa Fluor® dyes etc.) or using immuno-enzymatic staining (with peroxidase-labelled secondary antibodies). A variety of secondary antibodies have been used successfully (from rabbit, goat, sheep and donkey) as well as modified versions of these antibodies (e.g F(ab')2 derivatives).

 

The combinations of cell and tissue types, secondary antibodies and lables is too great for us to provide you with a synoptic overview here. However, we have compiled a database of publications, where you can search interactively. We also provide you with some additional information to common questions about the methods that have been used.