Paraffin-embedded 5-μm TMA sections were deparaffinized in xylene and rehydrated via soaking in decreasing percentages of ethanol solutions (100% twice, 90% and 70% once). Sections were heated in 0.1 M citrate for antigen retrieval, treated with 3% H2O2 to block endogenous peroxidase activity, and incubated overnight at 4°C with anti-OGT (1:50; ProteinTech, Chicago, IL), anti-MGEA5 (1:100; ProteinTech), or anti-O-GlcNAc (1:200; Thermo Fisher Scientific, Waltham, USA) primary antibodies. After washing in phosphate-buffer saline (PBS), sections were incubated with peroxidase-labeled secondary antibody for 1 h at room temperature. Sections were then incubated with diaminobenzidine, washed, and counterstained with hematoxylin. All stains were examined by pathologists and semi-quantitatively scored as follows: 0 (no staining), 1 (weakly positive), 2 (moderately positive), and 3 (strongly positive); percentage scores were 0 (0%), 1 (≤10%), 2 (11–50%), and 3 (51–100%). The IHC score for each specimen represents the intensity score multiplied by the percentage score, which ranged from 0–9.
Anti ogt
Manufactured by Proteintech
Sourced in United States
Anti-OGT is a laboratory reagent designed for the detection and quantification of O-GlcNAc transferase (OGT) in biological samples. OGT is an enzyme that catalyzes the addition of O-linked N-acetylglucosamine (O-GlcNAc) to serine and threonine residues of target proteins. Anti-OGT can be used in various research applications to study the role of OGT in cellular processes.
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Lab products found in correlation
Anti mgea5, by Proteintech (1 mentions)
Anti o glcnac, by Thermo Fisher Scientific (1 mentions)
Ecl plus kit, by GE Healthcare (1 mentions)
Western ip lysis buffer, by Beyotime (1 mentions)
Protein a g magnetic beads, by Selleck Chemicals (1 mentions)
Anti mouse igm hrp, by Abcam (1 mentions)
Anti ha magnetic beads, by Selleck Chemicals (1 mentions)
2 protocols using anti ogt
1
Immunohistochemical Analysis of OGT and MGEA5
2
Immunoblotting and Co-immunoprecipitation Protocols
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For immunoblots/lectinblots, the proteins were resolved on SDS-polyacrylamide gel electrophoresis gels followed by standard immunoblots. The primary antibodies used were anti-O-GlcNAc CTD110.6 (BioLegend, #838004), anti-OGT (Proteintech, #11576-2-AP), anti-GAPDH (CST, #5174), anti-HA (CST, #3724), anti-MTA1 (CST, #5646), anti-CHD4 (CST, #12011), anti-MBD3 (CST, #99169) and anti-HDAC1 (CST, # 34589). Lectin sWGA (Vector Laboratories, #B-1025S) was used for lectin blotting. The appropriate secondary antibody used were anti-mouse IgG-HRP (CST, # 7076), anti-rabbit IgG-HRP (CST, #7074), anti-mouse IgM-HRP (Abcam, #ab97230), and the signals were detected by the ECL Plus kit (GE Healthcare). All blots are representative of at least two independent experiments. For co-immunoprecipitation, cells were harvested and lysed with western/IP lysis buffer (Beyotime, #P0013). Immunoprecipitates were washed five times and then subjected to immunoblotting analysis. The antibodies and beads used for IP were anti-MTA1 (CST, #5646), anti-HA-magnetic beads (Bimake, #B26201) and protein A/G-magnetic beads (Bimake, #B23201).
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