Skip to main content
. 2014 Feb 26;289(16):11132–11142. doi: 10.1074/jbc.M113.492512

FIGURE 2.

FIGURE 2.

Detection of GlcNAc-terminated, complex N-glycans by other anti-O-GlcNAc antibodies. A, MFNG-AP affinity-purified from Lec8/M conditioned medium (upper) or Lec8/M whole cell lysate (lower) was separated in a 10% SDS-polyacrylamide gel, transferred to PVDF membranes, and incubated with anti-O-GlcNAc antibodies (top and bottom), or anti-human PLAP (L-19) antibody (middle), for 1 h at room temperature. O-GlcNAcylated proteins were detected with CTD110.6 (0.4 μg/ml) or 10D8 (0.2 μg/ml) and anti-mouse IgM antibody-HRP (1.6 μg/ml), RL2 (3 μg/ml), HGAC85 (4 μg/ml), #3 (0.86 μg/ml), #10 (0.59 μg/ml), or #14 (0.97 μg/ml) and anti-mouse IgG-HRP (0.08 μg/ml). Representative results are from two preparations. B, mAbs #3 (8.6 μg/ml) and #14 (9.7 μg/ml) were incubated at room temperature for 1 h followed by anti-mouse IgG-HRP (0.08 μg/ml). IP, immunoprecipitate; IB, immunoblot.