O-GlcNAc Monoclonal Antibody (RL2), Alexa Fluor™ 488, eBioscience™, Invitrogen™

Description

• Description: This RL2 monoclonal antibody recognizes proteins with O-N-acetylglucosamine (O-GlcNAc) glycosylation
• It was originally developed by immunizing mice with the rat liver nuclear envelopes containing nuclear pore complexes
• The RL2 clone has been successfully used in Western blot, immunofluorescence, immunoprecipitation, and flow cytometry in a wide variety of mammalian cells
• Applications Reported: This RL2 antibody has been reported for use in intracellular staining followed by flow cytometric analysis
• Applications Tested: This RL2 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol
• Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online
• This may be used at 5 μL (0.5 μg) per test
• A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• Excitation: 488 nm; Emission: 519 nm; Laser: Blue Laser O-linked N-acetylglucosamine (O-GlcNAc) is a posttranslational modification characterized by the attachment of N-acetylglucosamine to specific serine or threonine residues
• Unlike other protein glycosylations, O-GlcNAc modifications occur within the nucleus and cytoplasm
• They are found on many cellular proteins, including nuclear pore, oncogene, cytoskeletal, heat shock, viral and transcription regulatory proteins
• O-GlcNAc glycosylations are thought to obscure phosphorylation sites, counteracting phosphorylation-dependent signaling pathways and protein interactions.