1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 5 µg/ml.
Eukaryotic cilia are assembled via intraflagellar transport (IFT) in which large protein particles are motored along ciliary microtubules. The IFT20 subunit of a particle is localized to the Golgi complex, plus the basal body and cilia. In living cells, fluorescently tagged IFT20 is highly dynamic and moves between the Golgi complex and the cilium as well as along ciliary microtubules. It has been suggested that IFT20 functions in the delivery of ciliary membrane proteins from the Golgi complex to the cilium. There are three different isoforms.
ab103693 stained WI 38 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab103693 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - IFT20 antibody (ab103693)
Anti-IFT20 antibody (ab103693) at 1/100 dilution + Mouse lung tissue lysates at 35 µg
ab103693, at a 1/50 dilution, staining IFT20 in formalin fixed, paraffin embedded Human hepatocarcinoma tissue by Immunohistochemistry, followed by peroxidase conjugated secondary antibody and DAB staining.