TABLE 2.

Characteristics of the most commonly used O-GlcNAc antibodies.

	Antigen	Specificity
CTD110.6 (Cell Signaling mAb #9875)	O-GlcNAc modified peptide: YSPTS*PS	Can recognize O-GlcNAc-Ser, or O-GlcNAc-Thr. Cross reacts with N-GlcNAc2-modified proteins (Isono, 2011) and terminal β-glycans on complex N-glycans (Tashima and Stanley, 2014). Off-target binding may be alleviated by using PNGase F to remove N-linked glycans, and mild on-blot β-elimination was used to remove O-glycans
RL2 (Millipore MABS157)	Nuclear Envelope (NE) fractions from the rat liver	Adding galactose to the O-GlcNAc moiety or removing O-GlcNAc from the proteins decreased RL2 recognition of these proteins. It no longer recognizes NE antigen once they are partially proteolyzed. (Holt et al., 1987; Snow et al., 1987; Zachara et al., 2011; and Reeves et al., 2014)
1F5.D6(14), 9D1.E4(10), and 18B10.C7(3) (Millipore)	CKII peptide GSTPVS(β-O-GlcNAc)SANM	Bind BSA-CGSTPVS(β-O-GlcNAc)SANM not BSA-CGSTPVSSANM Inhibited by GSTPVS(β-O-GlcNAc)SANM, but not GSTPVSSANM or β-O-GlcNAc-Ser Binding to glycopeptide by 1F5.D6(14) (but not 9D1.E4(10) and 18B10.C7(3) only marginally) can also be inhibited by VS(β-O-GlcNAc)S and PVS(β-O-GlcNAc)SA