Western blot was performed on whole cell lysates from mouse embryonic stem cells (E14Tg2a) with Mll4 polyclonal antibody (Cat # PAB14067), diluted 1 : 500 in PBS-Tween containing 5% skimmed milk. Cells homozygous for the targeted mll4 allele (-/-, Lane 2 and 3) show a complete loss of Mll4 protein, whereas the Mll4 protein was detected in flp-recombined ES cells (F/F, Lane 4) and heterozygously targeted ES cells (+/-, Lane 1).
ELISA was performed using a serial dilution of Mll4 polyclonal antibody (Cat # PAB14067). The wells were coated with the peptides used for immunisation of the rabbit. By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1 : 15,000.