Bi mecy 0100

The antibodies to 5-methyl-cytosine (5mC) (Eurogentec, BI-MECY-0100), H3K9me3 (Abcam, ab8898), H3K27ac (Abcam, ab4729), ZFP57 (Abcam, ab45341), SETDB1 (Santa Cruz Biotechnology, sc-66884), and RNA Pol II (Santa Cruz Biotechnology, sc-899) are commercially available. Antibodies against AFF3 (Ab#1, antigen, a mixture of two peptides from amino acids 124–237 and amino acids 816–982 of human AFF3) were generated in our laboratory and have been described previously (Luo et al. 2012a (link)). A second antibody against AFF3 (Ab#2, antigen, human AFF3 amino acids 816–982) was also generated in our laboratory. Antigens were expressed as His tag fusion proteins in pET-16b, purified on NTA-agarose according to Qiagen's protocol, and sent to Pocono Rabbit Farm and Laboratory for immunization into rabbits.

The antibodies to 5-methyl-cytosine (5mC) (Eurogentec, BI-MECY-0100), KAP1 (Abcam, ab10484), and SETDB1 (Proteintech, 11231-1-AP) were purchased. Antibodies to AFF3 and AFF4 were described previously (Luo et al., 2012 (link)). A fragment of human AF9 (amino acids 406–498) was expressed as a His-tag fusion protein in pET-16b, purified on NTA-agarose according to Qiagen’s protocol and by HPLC, and then sent to Genscript for immunization into rabbits. Whole-cell lysates for western blots were prepared by lysing cells in SDS-PAGE loading buffer.