Product Specifications for Recombinant Human B3GALNT2 Protein, CF
Chinese Hamster Ovary cell line, CHO-derived human B3GALNT2 protein Gly35-Arg500, with C-terminal 6x His tag
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
55-63 kDa, reducing conditions
Measured by its ability to transfer GalNAc from UDP-GalNAc to benzyl-GlcNAc. The specific activity is >10 pmol/min/ μg, as measured under the described conditions
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
6 months from date of receipt, -20 to -70 °C as supplied.
3 months, -20 to -70 °C under sterile conditions after opening.
B3GALNT2 was first identified in 2004 through a BLAST search (1). The expressed B3GALNT2 strictly transferred GalNAc to terminal GlcNAc residue and synthesized a unique carbohydrate structure GalNAc-beta 1,3-GlcNAc, which was not known in humans or other mammals previously. The related native structure was only found later in a study of congenital muscular dystrophies (CMD), a heterogeneous group of inherited diseases characterized by the onset of muscle weakness at birth or within 2 years of life. A common subgroup within the CMDs are the dystroglycanopathies, characterized by reduced functional glycosylation of alpha -dystroglycan ( alpha -DG), a peripheral membrane protein that binds to several extracellular matrix components. Proper glycosylation of alpha -DG is critical to maintain structural integrity and force transmission between the cytoskeleton and the extracellular matrix for efficient signal transduction. Deleterious mutation in gene of B3GALNT2 was found in some of the patients of dystroglycanopathies (2). The complete structure of B3GALNT2 product was then found as an O-mannosyl trisaccharide GalNAc-beta 1,3-GlcNAc-beta 1,4-Man (3) with the mannose phosphorylated at the 6-position (4). The enzymatic activity of recombinant human B3GALNT2 was determined using a phosphatase-coupled assay (5) using GlcNAc beta 1-O-benzyl as acceptor substrate. Recombinant B3GALNT2 is also found to be highly active and specific towards O-GlcNAc and therefore is an ideal enzyme for the detection and labeling of O-GlcNAc (6). O-GlcNAc is a single GlcNAc residue attached to serine/threonine residues on nuclear and cytosolic proteins with regulatory roles from transcription, translation, cell signaling to cell cycle regulation (7).
Hiruma, t, et al. (2004) J. Biol. Chem. 279:14087.
Stevens, E. et al. (2013) Am. J. Hum. Genet. 92:354.
Yoshida-Moriguchi, T. et al. (2013) Science 341:6148.
Yoshida-Moriguchi, T. et al. (2010) Science 327:88.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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