All lanes : Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594) antibody (ab61791) at 1/1000 dilutionLane 1 : HeLa whole cell lysateLane 2 : HeLa whole cell lysate - overexpressing EphB2Lane 3 : HeLa whole cell lysate - overexpressinf EphB2-kinase dead (K661R)Lysates/proteins at 100000 cells per lane.SecondaryHRP-conjugated goat anti-rabbit IgG monoclonal at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ab61791 staining Eph receptor B1 + Eph receptor B2 (phospho Y594) in HeLa cells (overexpressing EphB2 and stimulated for 20 minutes with preclustered efnb1-Fc chimeric protein) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 1% BSA + 0.1% Triton X-100) for 2 hour at 25°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody. Red - anti-EphB2 (goat), Grey - DAPI, Green - ab61791.See Abreview
All lanes : Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594) antibody (ab61791) at 1/500 dilutionLane 1 : Extracts from HepG2 cellsLane 2 : Extracts from HepG2 cells with the immunising phosphopeptide
Immunofluorescence analysis of HuvEc cells, using ab61791 at 1/500-1/1000 dilution with (+) or without (-) immunising phosphopeptide.