Summary: haemagglutinin antibodies, and ectopically expressed full-length (fl) Alk (Twist-Gal4;
UAS-Alkfl
) was immunoprecipitated with anti-Alk antibodies.
ELISA and immunofluorescence analysis
ELISA was carried out according to standard protocols. Microtitre plate wells were coated
with Drosophila Alk monoclonal antibody 123 (ref. 1) for the sandwich ELISA, and with
purified recombinant His­Jeb protein for the direct ELISA. For immunofluorescence
staining, COS7 cells were transfected with either pcDNA3 (as control), pcDNA3-Alkfl
,
pcDNA3-AlkE
, or pcDNA3-AlkA
(where superscipt E and A indicated extracellular and
activated, respecitvely), and purified recombinant His­Jeb protein was added (to a final
concentration of 1 mg ml21
) for 1 h before staining, as described previously21
. Analysis was
carried out by confocal laser scanning microscopy (Leica).
In situ hybridization
In situ hybridization of whole-mount embryos was done with digoxigenin-labelled duf
RNA as probe and followed by antibody staining as described22

Source: Arnold, Jonathan - Nanoscale Science and Engineering Center & Department of Genetics, University of Georgia

Collections: Biotechnology