Why is DTT (dithiothreitol) added to the assay buffer in the assay described for Recombinant Human Glucosylceramidase/GBA Protein, CF, Catalog # 7410-GHB?

Name: Recombinant Human Glucosylceramidase/GBA Protein, CF
Brand: R&D Systems
SKU: 7410-GHB

The 5 mM DTT is added to the assay buffer in the assay protocol for catalog # 7410-GHB to prevent interchain disulfide bond formation that can cause aggregation of the protein and loss of enzyme activity over time.

Recombinant Human Glucosylceramidase/GBA Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 7410-GHB

Citations (9)

Product Datasheet / COA / SDS

Catalog #

Availability

Size / Price

Qty

Setting the standard in quality research reagents

Key Product Details

Source:	CHO
Accession #:	P04062.3
Applications:	Enzyme Activity

View Full Technical Details for Recombinant Human Glucosylceramidase/GBA Protein, CF

Complete Your Research

Product Specifications for Recombinant Human Glucosylceramidase/GBA Protein, CF

Source	Chinese Hamster Ovary cell line, CHO-derived human Glucosylceramidase/GBA protein Ala40-Gln536, with a C-terminal 6-His tag
Purity	>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Ala40
Predicted Molecular Mass	56 kDa
SDS-PAGE	60-75 kDa, reducing conditions
Activity	Measured by its ability to hydrolyze 4-methylumbelliferyl-beta -D-glucopyranoside. The specific activity is >200 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation and Storage

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.

7410-GHB

Formulation	Supplied as a 0.2 μm filtered solution in Sodium Citrate.
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.

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: Glucosylceramidase/GBA

Glucosylceramidase is a lysosomal enzyme that cleaves the beta-glucosidic linkage of glucosylceramide (1, 2), an intermediate in glycolipid metabolism. The mature enzyme has 497 amino acids with a molecular weight of 62 kDa (3). Glycosylation occurs at four of five N-glycosylation sites and is essential for the trafficking and activity of the enzyme (4). The enzyme is activated in lysosomes by saposin C, although the mechanism of activation is not well understood (5). Defects in Glucosylceramidase are the cause of Gaucher disease, also known as glucocerebrosidase deficiency (6). Gaucher disease is the most prevalent lysosomal storage disease, characterized by accumulation of glucosylceramide in the reticulo-endothelial system. Symptoms of Gaucher disease may include enlarged spleen and liver, liver malfunction, skeletal disorders and bone lesions, severe neurologic complications, swelling of lymph nodes, anemia, low blood platelets and yellow fatty deposits on the white of the eye (7). Currently, enzyme replacement therapy is used to treat patients with the disease (8, 9).

Item	Value
References	Sorge, J. et al. (1985) Proc. Natl. Acad. Sci. USA 82:7289. Ginns, E. I. et al. (1984) Biochem. Biophyl. Res. Commun. 123:574. Horowitz, M. et al. (1989) Genomics 4:87. Grace, M.E. et al. (1994) J. Biol. Chem. 269:2283. Bruhn, h. (2005) Biochem. J. 389:249. Liou, B. et al. (2006) J. Biol. Chem. 281:4242. Grabowski, G.A. (2008). Lancet 372:1263. Zheng, W. et al. (2007) Proc. Natl. Acad. Sci. USA 104:13192. Beutler, E. and Gelbart, T. (1996) Hum. Mutat. 8:207.
Entrez Gene IDs	2629 (Human); 14466 (Mouse)
Alternate Names	Acid beta-glucosidase, Alglucerase, D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45, GBA, GBA1, GC, GCB, GLUC, Glucosylceramidase, Imiglucerase, beta-glucocerebrosidase, glucosidase, beta, acid, glucosidase, beta; acid (includes glucosylceramidase), lysosomal glucocerebrosidase

Citations for Recombinant Human Glucosylceramidase/GBA Protein, CF (9)

Showing 1 - 9 of 9 citations Showing All

Filter By:

Search Title:

Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human Glucosylceramidase/GBA Protein, CF include:

Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

MC Babcock et al. (2021), Substrate reduction therapy for Krabbe disease and metachromatic leukodystrophy using a novel ceramide galactosyltransferase inhibitor Scientific Reports, 11(1):14486.
PMID: 34262084
Species: Human

Applications: Cell Culture

A Brown et al. (2021), Recapture Lysosomal Enzyme Deficiency via Targeted Gene Disruption in the Human Near-Haploid Cell Line HAP1 Genes, 12(7).
PMID: 34356092
Species: Human
Sample Types: CSF
Applications: Cell Culture

L Oftedal et al. (2020), Validation and assessment of preanalytical factors of a fluorometric in vitro assay for glucocerebrosidase activity in human cerebrospinal fluid Scientific Reports, 10(1):22098.
PMID: 33328543
Species: Mouse
Sample Types: Tissue Homogenates

H Li et al. (2019), Mitochondrial dysfunction and mitophagy defect triggered by heterozygous GBA mutations Autophagy, 15(1):113-130.
PMID: 30160596
Species: N/A
Sample Types: Protein
Applications: Enzyme Assay

MA Do et al. (2019), Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles Sci Rep, 9(1):17274.
PMID: 31754156
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay

SP Yun et al. (2018), ?-Synuclein accumulation and GBA deficiency due to L444P GBA mutation contributes to MPTP-induced parkinsonism Mol Neurodegener, 13(1):1.
PMID: 29310663
Species: Human
Sample Types: Fluorescent Probe
Applications: Bioassay

RA Ashmus et al. (2018), Fluorescence-Quenched Substrates for Quantitative Live Cell Imaging of Glucocerebrosidase Activity Meth. Enzymol., 598(0):199-215.
PMID: 29306435
Species: Human
Sample Types: Tissue Homogenates
Applications: Bioassay

Choi S et al. (2015), Lysosomal Enzyme Glucocerebrosidase Protects against Abeta1-42 Oligomer-Induced Neurotoxicity. PLoS ONE, 10(12):e0143854.
PMID: 26629917
Species: Human
Sample Types: Tissue Homogenates
Applications: Enzyme Assay

Berger et al. (2015), Tool compounds robustly increase turnover of an artificial substrate by glucocerebrosidase in human brain lysates. PLoS ONE, 10(3):e0119141.
PMID: 25763858

There are no citations that match your criteria.

Showing 1 - 9 of 9 citations Showing All

FAQs for Recombinant Human Glucosylceramidase/GBA Protein, CF

Why is DTT (dithiothreitol) added to the assay buffer in the assay described for Recombinant Human Glucosylceramidase/GBA Protein, CF, Catalog # 7410-GHB?

The 5 mM DTT is added to the assay buffer in the assay protocol for catalog # 7410-GHB to prevent interchain disulfide bond formation that can cause aggregation of the protein and loss of enzyme activity over time.