Enzymes

PNGase F (CarboClip)

CarboClip is a recombinant peptide-N-Glycosidase F (PNGase F), from Elizabethkingia sp, overexpressed in E. coli that contains a hexahistidine tag in its C-terminus. It cleaves the glycosidic bond between the reducing end GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides on N-linked glycoproteins.
CarboClip is provided glycerol-free as a lyophilized powder containing Tris-HCl, NaCl, and Na₂EDTA.

Applications

• N-glycan analysis of antibodies and glycoproteins,
• Deglycosylation of membrane proteins on cells, viruses and exosomes
• Removal of N-Linked Glycans for Native Protein Mass Spectrometry

Activity:
After reconstitution in 150 μL of UltraPure H₂O, 1 μl of CarboClip can deglycosylate > 95% of ribonuclease B (1 mg) in less than 20 minutes. Activity is calculated by SDS-PAGE as by densitometry.

Storage and Stability:
Lyophilized powder stored at -80 °C and -20 °C is stable for more than 1 year, at room temperature stable for more than 10 days. Reconstituted protein should be stored at 4 °C and is stable for 2-3 months.

B3 Galactosyl Transferase

N-acetyl-beta-D-glucosaminide beta-(1,3)-galactosyltransferase Size: 4 mU

Introduction:
B3GalT is a recombinant N-acetyl-beta-D-glucosaminide beta-(1,3)-galactosyltransferase. The enzyme transfers galactose from UDP-galactose to the N-acetylglucosamine or N-acetylgalactosamine termini of acceptor molecules such as GalNAc, GlcNAc, Lacto-N-triose, Globotetrose, Isoglobotetrose and GalNAcβ1,3Galβ1,4Glc. Besides, B3GalT has been used to produce the Human Milk Oligosaccharide (HMO) lacto‐N‐tetraose (LNT) and N-Acetyllactosamine (LacNAc) Type 1 O-glycans.
B3Gal Transferase can be used for synthesis of the precursor of H type 1 antigen of ABO histo-blood group. The H type 1 antigen in itself is the precursor of the following type 1 antigens: antigens A, antigen B, Lewis a (Le a ), Lewis b (Le b ) and Sialyl Lewis a (sLe a).

Source:
E. coli cells with a cloned bacterial B3GalT gene.

Storage Buffer:
The enzyme is provided in solution containing Tris buffer, NaCl and 10% glycerol

Activity:
The activity per µL is 40 µUnits/µL or 248 pmol/min/μL

Unit Definition:
One milliunit will transfer 1.0 nanomole of galactose from UDP-galactose to 4-Nitrophenyl N-acetyl-β-D-glucosaminide per min at pH 7.5 at 37 °C.

Storage:
Store at -20 °C >1 year. Avoid repeated freeze-thaw cycles, which decrease protein stability.