The stability of this material in solution has not been evaluated. However, product G4155, is a β-galactosidase suspension in 50% glycerol, 5 mM Tris buffer salts, 5 mM magnesium chloride, 0.5 mM DTT, 0.5 mM mercaptoethanol. pH 7.4. This item is stored at -20°C and is stable, when stored properly, for at least 1 year. Similar stability is expected for prepared solutions of G5635, when stored in aliquots. Avoid repeated freeze/thaw cycles.
G5635
β-Galactosidase from Escherichia coli
β-Galactosidase from Escherichia coli
Grade VIII, lyophilized powder, ≥500 units/mg protein
Synonym(s):
β-D-Galactoside galactohydrolase, Lactase
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1000 UNITS
$118.00
3000 UNITS
$266.00
5000 UNITS
$423.00
15000 UNITS
$1,016.50
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1000 UNITS
$118.00
3000 UNITS
$266.00
5000 UNITS
$423.00
15000 UNITS
$1,016.50
About This Item
Recommended Products
type
Grade VIII
Quality Level
form
lyophilized powder
specific activity
≥500 units/mg protein
mol wt
465 kDa
does not contain
BSA as extender
composition
Protein, ≥60% biuret
storage temp.
−20°C
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General description
β-Galactosidase is a tetramer consisting of four equal subunits of 135,000 each. It is a sulfhydryl containing enzyme, with 19 cysteine residues per subunit.[1]
Application
β-Galactosidase is conjugated to an antibody which specifically recognizes a target molecule (enzyme immunoassay or EIA).[2] β-Galactosidase is also used as a reporter enzyme to monitor the level of gene expression of a promoter.[3] It may be used as a positive control protein with anti-β-galactosidase antibodies.
Biochem/physiol Actions
β-galactosidase cleaves lactose into its monosaccharide components, glucose and galactose. It also catalyses the transglycosylation of glucose into allolactose, the inducer of β-galactosidase, in a feedback loop.
Physical properties
Tetramer molecular weight 465 kDa (subunits 116.3 kDa each)
Unit Definition
One unit will hydrolyze 1.0 μmole of o-nitrophenyl β-D-galactoside to o-nitrophenol and D-galactose per min at pH 7.3 at 37 °C.
Physical form
Contains Tris buffer salts and magnesium chloride
inhibitor
Product No.
Description
Pricing
substrate
Product No.
Description
Pricing
Storage Class
11 - Combustible Solids
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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Alan Merk et al.
IUCrJ, 7(Pt 4), 639-643 (2020-07-23)
We report the determination of the structure of Escherichia coli β-galactosidase at a resolution of ∼1.8 Å using data collected on a 200 kV CRYO ARM microscope equipped with a K3 direct electron detector. The data were collected in a single 24 h
Assays for bacterial mucin-desulfating sulfatases.
A M Roberton et al.
Methods in molecular biology (Clifton, N.J.), 125, 417-426 (2000-05-23)
K Kato et al.
Journal of immunology (Baltimore, Md. : 1950), 116(6), 1554-1560 (1976-06-01)
1. A method for the conjugation of the Fab' fragment of rabbit IgG with beta-D-galactosidase from Escherichia coli is described. The method consists of two main steps: treatment of the Fab' fragments containing sulfhydryl groups with excess N,N'-o-phenylenedimaleimide, to introduce
Raimond B G Ravelli et al.
Nature communications, 11(1), 2563-2563 (2020-05-24)
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Articles
For use as a marker in SDS-PAGE; Albumin from chicken egg white, For use as a marker in SDS-PAGE; L-Lactic Dehydrogenase from rabbit muscle, Type XI, lyophilized powder, 600-1,200 units/mg protein
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Can I store my enzyme in 50 mM Tris-HCl, pH 7.3, 5-10 mM β-mercaptoethanol, and 10 mM MgCl₂ and 50% glycerol for long-term preservation at -20°C for several months?
1 answer-
Helpful?
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Can this product be stores dissolved in PBS at -20 C?
1 answer-
The solution stability and storage of this product in PBS have not been determined. However, according to the Product Datasheet, solutions of this enzyme should be prepared fresh. Alternatively, a prepared solution of this enzyme is available as product G4155. This solution is stored at -20°C and contains 50% glycerol, 5 mM Tris buffer salts, 5 mM magnesium chloride, 0.5 mM DTT, and 0.5 mM mercaptoethanol at pH 7.4. Please see the links below for additional information.
G5635 Datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/211/865/g5635pis.pdfG4155 B-Galactosidase aqeuous suspension:
https://www.sigmaaldrich.com/product/sigma/g4155Helpful?
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Should b-gal solution be prepared in 50 mM Tris HCl, 10 mM beta-mercaptoethanol and 10 mM MgCl2, as advised in PIS? Because these components were in buffer prior to lyophilization (10 mM Tris HCl, 10 mM MgCl2, and 1 mM beta-mercaptoethanol). Thanks
1 answer-
For immediate use and analysis, Product G5635 can be reconstituted in 100 mM Sodium Phosphate Buffer, pH 7.3 as shown in the enzymatic assay:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/332/599/g5635enz.pdfHowever, oxidation may be minimized and stability may be improved by preparing solutions in 50 mM Tris HCl, pH 7.3, 5-10 mM BME, and 10 mM MgCl2.
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