Current Project(s) :

siRNA cell array assays and characterisation of antibodies raised against novel cDNA products.

siRNA cell arrays are a convenient format for down-regulating hundreds of individual proteins in parallel. siRNAs are spotted on to chambered coverglass slides and cells are overlaid. The cells locally take up the siRNA designed against the specific target, including a number of novel proteins identified in the ‘gfp-cdna’ project, resulting in a defined cell population down-regulated for the target protein. Assays to determine any effects on membrane transport processes or organelle morphology are then performed. This allows us to identify novel proteins with a putative role in membrane transport.

So far we have identified a large number of novel molecules, which localise to compartments of the secretory pathway and potentially play a role in its function. For those, which appear to be most interesting, we raise polyclonal antibodies against either the whole protein (when purified), or against peptide sequences within the protein. Each of these antibodies must be purified and tested, for immunofluorescence and western blotting.