Immunofluorescence analysis of OGT was performed using 70% confluent log phase Panc-1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with OGT (HGAC85) Mouse Monoclonal Antibody (MA1076) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||All|
|Published species reactivity||Rabbit, Rat, Nematode, Hamster, Sheep, Human, Not Applicable|
|Host / Isotype||Mouse / IgG3|
|Immunogen||Heat killed, pepsin treated group A streptococci.|
|Storage buffer||ascites diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||1:20|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:250-1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunocytochemistry (ICC)||See 1 publications below|
|Western Blot (WB)||See 2 publications below|
|ChIP assay (ChIP)||See 1 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
|Immunoprecipitation (IP)||See 2 publications below|
|ELISA (ELISA)||See 1 publications below|
MA1-076 recognizes beta-1,3 linked O-linked N-acetylglucosamine (O-GlcNAc) residues of streptococcal group A carbohydrate as well as O-GlcNAc glycosylated proteins.
MA1-076 has been successfully used in Western blot, immunofluorescence, immunoprecipitation and ELISA procedures. By Western blot, this antibody detects several proteins representing immunoprecipitated O-GlcNAc glycoproteins. Immunofluorescence staining of O-GlcNAc in cells results in labeling of the nuclear envelope and pores, nucleolus, and cytoplasm. This staining pattern is consistent with other methods of detecting O-GlcNAc moieties.
The MA1-076 immunogen is heat killed, pepsin treated group A streptococci.
DO NOT USE WITH DILUENTS CONTAINING GLYCOSYLATED PROTEINS.
Many cellular proteins, including nuclear pore, oncogene, cytoskeletal, heat shock, viral and transcription regulatory proteins contain single O-linked N-acetylglucosamine (O-GlcNAc) residues attached to serine or threonine residues. It has been observed that O-GlcNAc glycosylated proteins tend to be under phosphorylated relative to unglycosylated proteins and that O-GlcNAc bearing proteins tend to be found in multimeric complexes. This has led to the suggestion that O-GlcNAc glycosylation may obscure phosphorylation sites and acts as a signaling mechanism or mediator of signaling.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Drosophila O-GlcNAcase Deletion Globally Perturbs Chromatin O-GlcNAcylation.
MA1-076 was used in immunocytochemistry to analyze global pertubing of chromatin O-GlcNAcylation after drosophila O-GlcNAcase deletion
|Akan I,Love DC,Harwood KR,Bond MR,Hanover JA||The Journal of biological chemistry (291:9906)||2016|
Antibodies that detect O-linked ß-D-N-acetylglucosamine on the extracellular domain of cell surface glycoproteins.
MA1-076 was used in western blot to charcaterize antibodies suitable for the dection of O-linked beta-D-N-acetylglucose on secreted proteins and the extracellular domain of membrane proteins
|Tashima Y,Stanley P||The Journal of biological chemistry (289:11132)||2014|
Investigations into charge heterogeneity of wool intermediate filament proteins.
MA1-076 was used in western blot to investigate the isoelectric charge status of wool intermediate filament proteins
|Paton LN,Gerrard JA,Bryson WG||Journal of proteomics (71:513)||2008|
Dynamic O-GlcNAc cycling at promoters of Caenorhabditis elegans genes regulating longevity, stress, and immunity.
MA1-076 was used in chimmunoprecipitation to determine the effects of GlcNAcylation on transcription in C. elegans genome
|Love DC,Ghosh S,Mondoux MA,Fukushige T,Wang P,Wilson MA,Iser WB,Wolkow CA,Krause MW,Hanover JA||Proceedings of the National Academy of Sciences of the United States of America (107:7413)||2010|
Myosin16b: The COOH-tail region directs localization to the nucleus and overexpression delays S-phase progression.
MA1-076 was used in immunohistochemistry to study the role of the myosin16b COOH-tail region in localization to the nucleus and S-phase progression
|Cameron RS,Liu C,Mixon AS,Pihkala JP,Rahn RJ,Cameron PL||Cell motility and the cytoskeleton (64:19)||2007|
Molecular alterations underlie nodal and paranodal degeneration in type 1 diabetic neuropathy and are prevented by C-peptide.
MA1-076 was used in immunoprecipitation to demonstrate that the progressive molecular aberrations are preventable by insulinomimetic C-peptide
|Sima AA,Zhang W,Li ZG,Murakawa Y,Pierson CR||Diabetes (53:1556)||2004|
Cytoplasmic O-glycosylation prevents cell surface transport of E-cadherin during apoptosis.
MA1-076 was used in immunoprecipitation to study the role of cytosolic O-glycosylation of E-cadherin in blocking its transport to the cell surface during apoptosis
|Zhu W,Leber B,Andrews DW||The EMBO journal (20:5999)||2001|
Monoclonal antibodies to streptococcal group A carbohydrate. I. A dominant idiotypic determinant is located on Vk.
MA1-076 was used in ELISA to characterize monoclonal antibodies against streptococcal group A carbohydrate
|Nahm MH,Clevinger BL,Davie JM||Journal of immunology (Baltimore, Md. : 1950) (129:1513)||1982|