OGT Antibody (YA257)

OGT Antibody (YA257) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to OGT.

OGT is an enzyme that catalyzes the transfer of a single N-acetylglucosamine from UDP-GlcNAc to serine or threonine residues in cytoplasmic and nuclear proteins, modifying them with a beta-linked N-acetylglucosamine (O-GlcNAc). It glycosylates a wide range of proteins, including histone H2B, AKT1, AMPK, ATG4B, CAPRIN1, EZH2, and others, regulating their cellular functions through crosstalk between glycosylation and phosphorylation or by affecting proteolytic processing. In muscle and adipocyte cells, OGT contributes to insulin resistance by glycosylating insulin signaling components and inhibiting AKT1 phosphorylation at 'Thr-308' (by similarity). Additionally, OGT regulates glycolysis by mediating the glycosylation of 6-phosphofructokinase PFKL, thereby inhibiting its activity. In chromatin structure, OGT plays a crucial role by mediating O-GlcNAcylation of histone H2B at 'Ser-112' and is recruited to CpG-rich transcription start sites of active genes via interaction with TET proteins (TET1, TET2, or TET3). The mitochondrial isoform (mOGT) exhibits cytotoxicity and induces apoptosis in various cell types, including the insulinoma cell line INS1.

Free

O15294

Predicted band size: 117 kDa; Observed band size: 117 kDa

Protein A affinity purified.

Nucleus. Cytoplasm. Membrane.

Non-conjugated

Unmodified

AB_3102382

Neuroscience

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:2,000; IF: 1:100; IHC-P: 1:200-1:1,000; FC: 1:1,000; IP: 1-2 μg/sample

• 
  Western blot analysis of extracts from Hela (lane 2(20μg) , NIH/3T3 (lane 3(20μg) , HEK293 (lane 4(20μg),using OGT Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of MCF-7 cells labeling OGT with OGT Antibody (HY-P80254)at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with OGT Antibody HY-P80254) at 1/500 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of HeLa cells labeling OGT with OGT Antibody (HY-P80254)at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with OGT Antibody (HY-P80254) at 1/500 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse testis tissue using OGT Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80254, 1/400) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse testis tissue using OGT Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80254, 1/400) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Flow cytometric analysis of 1X10^6 Hela cells labeling OGT Antibody(HY-P80254, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/1000 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

WB, IHC-P, ICC/IF, IP, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human OGT.AA range:997-1046.