Western blot analysis of O-GlcNAc was performed by loading 10µg of HEK293T or C2C12 whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing O-GlcNAc (Product #MA1-039) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were then washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product #31430) at a dilution of 1:15000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Dura (Product #34075). The image is a Western blot with antibody MA1-039 with untreated (lanes 1 and 3) versus treated (lanes 2 and 4) for O-GlcNAc enrichment.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||SANM-conjugated linear peptide corresponding to human O-GluNAc.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||5 µg|
|Western Blot (WB)||1:1000|
* 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.
MA1-039 has been successfully used in Western blot and immunoprecipitation applications with human samples. In Western blot, MA1-039 detects proteins with O-GlcNAc post-translational modification, and therefore will detect multiple specific bands.
O-linked N-acetylglucosamine (O-GlcNAc), is a member of the O-GlcNAc transferase family. O-GlcNAc is a posttranslational modification characterized by the attachment of N-acetylglucosamine to specific serine/threonine hydroxyl groups. It plays an important role in nutrient sensing, gene expression, and protein degradation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.