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O-GlcNAcase Assay Kit



(OGA, NAG, MGEA5 Assay Kit)



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 (J. Clin. Path. 13 353, 1960). 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.

#O-GlcNAcase​


Kit Components

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

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

OGA Control Solution: 10 ml, store at 4ºC


SDS:

p-nitrophenyl-b-N-acetyl-glucosaminide, TX-100, sodium acetate, acetic acid


Related Kits:

Nagalase assay


Citation:

Involvement of NDPK-B in glucose metabolism-mediated endothelial damage via activation of the hexosamine biosynthesis pathway and suppression of O-GlcNAcase activity.

Cells 9:2324, 2020


Laminar Flow on Endothelial Cells Suppresses eNOS O-GlcNAcylation to Promote eNOS Activity.

Circulation Research 129:1054, 2021