WB
Western blot analysis of OGT on different lysates with Rabbit anti-OGT antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: MCF7 cell lysate (20 µg/Lane) Lane 3: HEK-293 cell lysate (20 µg/Lane) Lane 4: NIH/3T3 cell lysate (20 µg/Lane) Lane 5: PC-12 cell lysate (20 µg/Lane) Lane 6: Mouse brain tissue lysate (40 µg/Lane) Lane 7: Rat brain tissue lysate (40 µg/Lane) Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-OGT antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. 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.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling OGT with Rabbit anti-OGT antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-OGT antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling OGT. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
OGT was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-OGT antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-OGT antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-OGT antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-OGT antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 59 seconds.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-OGT antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. 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.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-OGT antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. 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.| Product Name | OGT Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human OGT aa 997-1,046 / 1,046. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:2000 IHC:1:200-1:1000 ICC:1:100 FC:1:1000 IP: 1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | OGT |
|---|---|
| Gene Synonyms | OGT1 HRNT1 MRX106 XLID106 HINCUT-1 O-GLCNAC |
| Gene Full Name | O-linked N-acetylglucosamine (GlcNAc) transferase |
| Gene Summary | This gene encodes a glycosyltransferase that catalyzes the addition of a single N-acetylglucosamine in O-glycosidic linkage to serine or threonine residues. Since both phosphorylation and glycosylation compete for similar serine or threonine residues, the two processes may compete for sites, or they may alter the substrate specificity of nearby sites by steric or electrostatic effects. The protein contains multiple tetratricopeptide repeats that are required for optimal recognition of substrates. Alternatively spliced transcript variants encoding distinct isoforms have been found for this gene. [provided by RefSeq, Oct 2009] |
| Alternative Names | FLJ23071 antibody GlcNAc transferase antibody HRNT1 antibody MGC22921 antibody O GlcNAc transferase p110 subunit antibody O GlcNAc transferase subunit p110 antibody O linked N acetylglucosamine (GlcNAc) transferase (UDP N acetylglucosamine:polypeptide N acetylglucosaminyl transferase) antibody O linked N acetylglucosamine (GlcNAc) transferase antibody O linked N acetylglucosamine transferase 110 kDa subunit antibody O-GlcNAc transferase subunit p110 antibody FLJ23071 antibody GlcNAc transferase antibody HRNT1 antibody MGC22921 antibody O GlcNAc transferase p110 subunit antibody O GlcNAc transferase subunit p110 antibody O linked N acetylglucosamine (GlcNAc) transferase (UDP N acetylglucosamine:polypeptide N acetylglucosaminyl transferase) antibody O linked N acetylglucosamine (GlcNAc) transferase antibody O linked N acetylglucosamine transferase 110 kDa subunit antibody O-GlcNAc transferase subunit p110 antibody O-linked N-acetylglucosamine transferase 110 kDa subunit antibody ogt antibody OGT1_HUMAN antibody UDP N acetylglucosamine peptide N acetylglucosaminyltransferase 110 kDa subunit antibody UDP N acetylglucosamine peptide N acetylglucosaminyltransferase GlcNAc transferase antibody UDP-N-acetylglucosamine--peptide N-acetylglucosaminyltransferase 110 kDa subunit antibody UDP-N-acetylglucosamine:polypeptide-N-acetylglucosaminyl transferase antibody Uridinediphospho N acetylglucosamine:polypeptide beta N acetylglucosaminyl transferase antibody |
| Molecular Weight(MW) | 117kDa |
| Cellular Localization | Nucleus. Cytoplasm. Membrane. |
WB
Western blot analysis of OGT on different lysates with Rabbit anti-OGT antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: MCF7 cell lysate (20 µg/Lane) Lane 3: HEK-293 cell lysate (20 µg/Lane) Lane 4: NIH/3T3 cell lysate (20 µg/Lane) Lane 5: PC-12 cell lysate (20 µg/Lane) Lane 6: Mouse brain tissue lysate (40 µg/Lane) Lane 7: Rat brain tissue lysate (40 µg/Lane) Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-OGT antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. 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.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling OGT with Rabbit anti-OGT antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-OGT antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling OGT. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
OGT was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-OGT antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-OGT antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-OGT antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-OGT antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 59 seconds.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-OGT antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-OGT antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. 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.| Application Notes | WB:1:2000 IHC:1:200-1:1000 ICC:1:100 FC:1:1000 IP: 1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
