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O-GlcNAcase (OGA, NAG or MGEA5) Assay Kit

Components:   

10x Cell Lysis Solution: 25 ml, store at 4ºC

OGA Assay Solution: 20 ml, store in aliquots at -20ºC

Product Description: In higher eukaryotes post-translational modification of cytoplasmic and nuclear proteins by O-linked
N-acetylglucosamine (O-GlcNAc) on serine and threonine residues is analogous to protein phosphorylation, serving to regulate protein function
and diverse cellular processes. The enzymes responsible for O-GlcNAc addition and removal are O-GlcNAc transferase (OGT) and
O-GlcNAcase (OGA), respectively. OGA is also known as b-N-acetylglucosaminidase (NAG) or MGEA5. Studies have shown that in addition to
their catalytic roles in protein modification, OGT and OGA possess multifunctional domains to target O-GlcNAc cycling to discrete intracellular
sites, thus influencing diverse kinase and phosphatase signaling activities. The OGA assay is based on the cleavage by OGA of the artificial
substrate p-nitrophenyl-beta-N-acetyl-glucosaminide to nitrophenol in an acidic buffer. Ionization of nitrophenol by NaOH produces a yellow
color exhibiting an absorption maximum at 405-415 nm, which allows for sensitive detection of OGA activity in crude cell/tissue extracts,
serum/plasma and urine. The assay is designed for the 96-well plate format, but can be scaled up if desired. Repeated freeze-thaw cycles of
the assay solution should be avoided. Use 1N NaOH as stop solution for the assay (not provided).