Product Specifications for Recombinant S. pyogenes Endo S2 His-tag Protein, CF
E. coli-derived s. pyogenes Endo-beta-N-acetylglucosaminidase S2/Endo S2 protein Glu37-Asp843, with a N-terminal Met & 6-His tag
>90%, 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
86-93 kDa, under reducing conditions.
Measured by its ability to digest Cy5-Labeled Glycan G2 >50% of Cy5-Labeled Glycan G2 (0.2 pmol) is digested by 0.2 μg of rSp. Endo-S2, as measured under the described conditions.
Scientific Data Examples for Recombinant S. pyogenes Endo S2 His-tag Protein, CF
Recombinant Sp. Endo S2 (Endo S2) Enzyme Activity Diagram
Endo S2 recognizes the conserved N-glycans on the Fc region of IgG by hydrolyzing the chitobiose core (at the beta -1,4 linkage between the two N-acetylglucosamines) of the N-glycans. Endo S2 leaves one GlcNAc residue attached the the asparagine of the peptide backbone. R1 and R2 can be oligosaccharide extensions containing Gal, GlcNAc, and Sialic Acid. R3 can be unmodified or Core-6 Fucose.
Recombinant S. pyogenes Endo S2 SDS-PAGE
1 μg/lane of rSp. Endo-S2 (Catalog # 10976-GH) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a band at 92 kDa.
Fluorescent Gel Mobility Shift caused by rSp. Endo S2.
Lane 1 contained substrate Cy5-labeled G2 GL302. In the presence of Endo S2, the glycan was digested to products and the smaller product Fuc-alpha,6-GlcNAc is observed.
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.
Streptococcus pyogenes is a leading Gram-positive bacterial pathogen that can abolish the effector functions of human immunoglobulin G (IgG) through deglycosylation (1). Upon infection, the pathogen secret two endoglycosidases, Endo S and Endo S2, that specifically deglycosylate the conserved N-glycans on the Fc region of IgG by hydrolyzing the chitobiose core (at the beta -1,4 linkage between the two N-acetylglucosamines) of the N-glycans (2, 3). Endo S and S2 cleavage leave one GlcNAc residue remaining attached to the asparagine residue on the peptide backbone. The enzymes are highly specifc to native IgG molecules (3), suggesting that the local conformation of IgG is required for the enzymatic recognition. In comparison, PNGase F from Flavobacterium meningosepticum completely removes glycans from glycoproteins and is more active on denatured glycoproteins. Cleavage of the glycans from IgG antibodies by Endo S and S2 result in conformation change of the antibodies thereby dramatically diminish the binding affinity to their receptors (4) and abolish their opsonizing functions (5, 6). By using fluorophore labeled N-Glycans as substrates, we found that Endo S2 has similar activity to Endo S towards non-galactosylated IgG glycan species including oligomannose and hybrid glycans but shows much high activity on galactosylated and sialylated IgG glycan species. We also found that both enzymes are highly active on core-6 fucosylated IgG glycans but with no activities on those with bisecting GlcNAc.
Nizet, V. (2007) J. Allergy Clin. Immunol. 120:13.
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