Home / Endo-beta-N-acetylglucosaminidase H/Endo H / Recombinant S. plicatus Endo H Protein, CF

Recombinant S. plicatus Endo H Protein, CF

Catalog #: 5224-GHB Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
5224-GHB-050
R&D Systems Recombinant Proteins and Enzymes
1 Image
Product Details
FAQs
Reviews
Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant S. plicatus Endo H Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<0.01 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to deglycosylate ribonuclease B under native conditions. The DC50 is <5 ng. The DC50 is defined as the amount of enzyme required to remove 50% of glycan on 1 μg of RNase B in 30 minutes at
37 °C.. Use of Recombinant S. plicatus Endo-beta -N-acetylglucosaminidase H/Endo H in the deglycoslyation of other substrates may require alternative conditions for optimal performance.
Source
E. coli-derived s. plicatus Endo-beta-N-acetylglucosaminidase H/Endo H protein
Val47-Pro313 with an N-terminal Met and 6-His tag
Accession #
P04067
N-terminal Sequence
Analysis
MHHHHHHVKQ
Predicted Molecular Mass
30 kDa
SDS-PAGE
29 kDa, reducing conditions

Product Datasheets

You must select a language.

x

5224-GHB

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. 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 could interfere.

5224-GHB

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and EDTA.
Shipping 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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Sodium Phosphate, pH 5.5
  • Recombinant S. plicatus Endo-beta -N-acetylglucosaminidase H/Endo H (rS. plicatus Endo H) (Catalog # 5224-GHB)
  • RNase B (Sigma, Catalog # R1153), 1 mg/mL stock in 10 mM Sodium Phosphate, pH 5.5
  • 4-20% SDS-PAGE gel
  • Reducing SDS-PAGE gel loading buffer
  • Silver Staining reagents
  1. Dilute Endo H to 2.5, 1.25, 0.625. 0.313, 0.156, 0.0781, and 0.0391 μg/mL in Assay Buffer.
  2. Dilute RNase B to 100 μg/mL in Assay Buffer.
  3. Combine 25 μL of Endo H at each dilution with 25 μL of 100 μg/mL RNase B. Include a control containing 25 μL Assay Buffer and 25 μL of 100 μg/mL RNase B.
  4. Incubate the reaction and control at 37 °C for 30 minutes.
  5. Combine 30 μL of  each reaction with 30 μL of reducing SDS-PAGE gel loading buffer. Heat samples at 100 °C for 3 to 5 minutes before loading to gel.
  6. Load 40 μL of sample per lane (1 μg of RNase B per lane) on a 15% gel.
  7. Stain gel with silver stain.
  8. Determine the 50% deglycosylation concentration (DC50) for Endo H by plotting % of RNase B deglycosylated vs. Endo H concentration with 4-PL fitting.
Per Lane:
  • rS. plicatus Endo H: 25, 12.5, 6.25, 3.13, 1.56, 0.781, and 0.391 ng
  • RNase B: 1 µg
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: Endo-beta-N-acetylglucosaminidase H/Endo H

N-glycans are commonly found on various glycoproteins. While peptide N-glycosidase from Flavobacterium meningosepticum (PNGase F) is widely used to release virtually all types of N-glycans under denaturing conditions, some bacterial endo-beta -N-acetylglucosaminidases, including Endo H, can be used under native conditions to specifically release particular types of N-glycans (1, 2). Because these glycosidases hydrolyze the chitobiose core of an N-glycan on a glycoprotein, the released glycan product will contain one GlcNAc residue at its reducing end with the other GlcNAc residue remaining attached to the glycoprotein. Endo H specifically releases oligomannose and hybrid but not complex type N-glycans from glycoproteins (3-5). It also remains active on core fucosylated N-glycans but not sulfated glycans (5).

References
  1. Maley, F. et al. (1989) Anal. Biochem. 180:195.
  2. Tarentino, A.L. et al. (1985) Biochemistry 24:4665.
  3. Hsieh, P. et al. (1983) J. Biol. Chem. 258:2555.
  4. Tarentino, A.L. et al. (1992) J. Biol. Chem. 267:3868.
  5. Trimble, R.B. and Tarentino, A.L. (1991) J. Biol. Chem. 266:1646.
Alternate Names
EndobetaNacetylglucosaminidase H; Endo-beta-N-acetylglucosaminidase H

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs

Reviews for Recombinant S. plicatus Endo H Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant S. plicatus Endo H Protein, CF and earn rewards!

Have you used Recombinant S. plicatus Endo H Protein, CF?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review