We have gone recombinant! Our most useful, widely used and best cited mouse monoclonal antibodies have now been cloned, sequenced and expressed In Vitro, meaning that they are now essentially immortal. Antibodies made from hybridoma lines may have problems due to loss of expression, mutation, infection or lab accidents. As a result some of the highest impact journals now prefer scientists to use recombinant antibodies if they available to optimize experimental reproducibility, see for example here and here. We have cloned, sequenced and expressed the top 20 EnCor mouse monoclonal antibodies and will market them in the recombinant form in the near future. We also generated recombinant forms of three of our most widely used antibodies now with rabbit IgG backbones to enhance their versatility. These are a rabbit monoclonal to NeuN/FOX3 RRMCA-1B7 and two rabbit monoclonals to c-FOS, RRMCA-2H2 and RRMCA-1B62. The RRMCA-1B7 antibody inherits the extensive data we have for the original mouse reagent including the epitope, kinetic properties, species cross reactivity and utility for WB, IF, ICC and IHC, making it the best characterized NeuN/FOX3 reagent available. The RRMCA-1B62 antibody, like the original MCA-1B62, was developed to be particularly insensitive to formalin fixation and so works well on formalin fixed paraffin embedded histological sections. The MCA-2H2 clone has become very widely used but is less robust on heavily fixed material for IHC. We have extensively tested all three antibodies and they function exactly like the originals. These novel rabbit monoclonal antibodies will be a particularly useful for ultra reliable application on mouse tissues where endogenous mouse IgG make technical problems.
We are well known for the quality of our antibodies and, using them, the resulting fluorescent images. A few years ago we took some sample stained microscope slides to the annual Society for Neuroscience Meeting and took them by various microscope vendors. It is fair to say that they were extremely impressed and so we have been supplying them to most major microscope companies for the last few years. So we have decided to make them generally available from our website. They can used to optimize the performance of fluorescence, confocal and multiphoton microscopes or for general teaching purposes. The slides use a collection of EnCor primary antibodies made in different species, typically mouse, rabbit, chicken and goat which are each specifically detected with appropriate species specific secondary reagents coupled to some combination of 488nm, 561nm, 594nm, 640nm and 750nm wavelengths. We also utilize the DNA fluorescent dye DAPI which emits at a 405nm wavelength and reveals cell nuclei. Here is an example of a video, now posted on Youtube, showing stained of HeLa cells with 5 channels here, in this case revealing microtubules with β-tubulin antibody (MCA-1B12), nucleoli with fibrillarin antibody (RPCA-Fib), mitochondria with HSP60 antibody (CPCA-HSP60), all from EnCor. We also revealed nuclear DNA with DAPI and microfilaments with fluorescent phalloidin. We decided to market similar preparations of 3T3 cells as shipping of human derived cells across national boundaries can be problematic. Such slides will be available shortly.
