All reagents and solvents were acquired from Fisher Scientific and were used as received unless otherwise stated. Electrophoresis and western blotting were performed by using Bio-Rad equipment. Primary antibodies were obtained from various sources: O-GlcNAc antibody from Biolegend, GAPDH antibody from R&D Systems, and PARP antibody from TACS. α-Crystallin was purchased from Sigma–Aldrich. Photomicrographs were acquired on a BZ-X700 Keyence Microscope, and all images were processed by using ImageJ software. Statistical analysis and graph generation was completed by using GraphPad Prism 7 software. Human brain tissue specimens were obtained from the Harvard Brain Tissue Resource Center. The human tissue array MC245a was purchased from US Biomax.
O glcnac antibody
Manufactured by BioLegend
The O-GlcNAc antibody is a tool for detecting the post-translational modification of O-linked N-acetylglucosamine (O-GlcNAc) on proteins. It is used to identify and quantify the presence of O-GlcNAc on target proteins in various biological samples.
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2 protocols using o glcnac antibody
1
Analyzing O-GlcNAcylation in Human Brain Tissue
2
Quantifying O-GlcNAcylated APP in Lipid Rafts
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To measure levels of O-GlcNAcylated APP in lipid raft (#4–6) and non-raft (#8–12) fractions, protein concentrations of each fraction were measured (Bio-Rad, Tokyo, Japan, #5000006). Equal amounts of proteins (400 μg) from lipid raft and non-lipid raft fractions were incubated with 4 μL of O-GlcNAc antibody (BioLegend, monoclonal, RRID: AB_2629520) or APP antibody (6E10, BioLegend, monoclonal, RRID: AB_2564652) for 2 h at room temperature, and then incubated with Protein G agarose Fast Flow (Millipore, #16266) for 16 h at 4 °C. Immunoprecipitated samples were then washed three times with PBS. Equal volumes of each fraction were loaded on a Western blot to detect O-GlcNAcylated APP.
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