| Description | Product DescriptionBeta Galactosidase from Streptococcus pneumoniae releases only β(1-4)- linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins. β(1-4) galactose is by far the most common linkage found in N-linked oligosaccharides. For other galactosidase Product DescriptionBeta Galactosidase from Streptococcus pneumoniae releases only β(1-4)- linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins. β(1-4) galactose is by far the most common linkage found in N-linked oligosaccharides. For other galactosidase linkages, ß(1-3,4,6)-Galactosidase from Bovine testes is recommended. The enzyme is as active on tetraantennary oligosaccharides as on disaccharides containing β(1-4)-linked galactose. Fucose linked to the penultimate N-acetylglucosamine will block cleavage of the galactose. Up to 100 υg of asialofetuin can be completely β(1-4)-degalactosylated in less than 1 hour using 3 mU of enzyme.Contentsß-(1-4) Galactosidase in 20 mM Tris-HCl, 25 mM NaCl (pH 7.5).Included with 20 µL and 60 µL pack sizes:5x Reaction Buffer 6.0 (250 mM sodium phosphate, pH 6.0).Molecular Weight ~250,000 daltonspH optimum 6.0, active over the range 5-7.The supplied buffer concentrate provides the optimal pH for enzyme activity with the standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity.Formulation The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl (pH 7.5).Specific ActivityOne unit of ß-(1-4)-galactosidase is defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 37°C pH 5 from p-nitrophenyl-beta-D-galactopyranoside.Specificity Non-reducing terminal ß(1-4)-Galactose. Number of antennae does not affect cleavage rate. Fucose linked to the penultimate N-acetylglucosamine will block cleavage of the galactose.Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.Purity ß(1-4)-Galactosidase is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours with 2 µl of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. The production host strain has been extensively tested and does not produce any detectable glycosidases.Directions for use 1. Add up to 100 µg of asialoglycoprotein or 1 nmol of oligosaccharide to tube. 2. Add deionized water to a total of 14 µl. 3. Add 4 µl of 5x Reaction Buffer 6.0. 4. Add 2 µl ß(1-4) Galactosidase. 5. Incubate at 37°C for 1 hour. For glycoproteins, cleavage may be monitored by SDS-PAGE if the size differential between native and de-galactosylated protein is sufficient for detection. Note: The optimum pH for cleavage of oligosaccharides is ~6... Read More | Inquire | Laccase is an enzyme, produced by ericoid mycorrhiza and ectomycorrhiza fungi. It belongs to the group of polyphenol oxidases. Laccase is also present in plants and bacteria.Laccase from Trametes versicolor has been used: to assess the use of four laccase-producing strains in waste water treatment Laccase is an enzyme, produced by ericoid mycorrhiza and ectomycorrhiza fungi. It belongs to the group of polyphenol oxidases. Laccase is also present in plants and bacteria.Laccase from Trametes versicolor has been used: to assess the use of four laccase-producing strains in waste water treatment in laccase assay in screening the lignolsSome of the enzymatic actions of laccase are associated with sporulation, detoxification, morphogenesis, melanin polymerization and it offers protection to spore coat. Laccase can catalyse a number of substrates including medicinal drugs and halogenated pesticides. It utilizes oxygen for its catalysis. For these reasons, it might be useful in the biological degradation of micropollutants in wastewater treatment. Laccase catalyzes the oxidation of phenol containing compounds, including lignin, through the reduction of oxygen to water. The presence of mediators will allow the oxidation of non-phenlic compounds as well. The primary function of laccase is to degrade lignin in fungi... Read More | Purity> 97 % by SDS-PAGE and HPLC analyses.FunctionReceptor for TNFSF2/TNF-alpha and homotrimeric TNFSF1/lymphotoxin-alpha. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation whichPurity> 97 % by SDS-PAGE and HPLC analyses.FunctionReceptor for TNFSF2/TNF-alpha and homotrimeric TNFSF1/lymphotoxin-alpha. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. Contributes to the induction of non-cytocidal TNF effects including anti-viral state and activation of the acid sphingomyelinase... Read More | TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP.TMB-D Blotting liquid ready-to-use substrate is a highly active and stable blotting substrate utilized for TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP.TMB-D Blotting liquid ready-to-use substrate is a highly active and stable blotting substrate utilized for measuring HRP probe activity. A stable blue precipitate is formed at the reaction site.The substrate does not contain NMP (1-methyl2-pyrrolidone) making it REACH Restricted Substances List Annex XVII compliant, while ensuring maximal safety during use, and minimal negative environmental impact.Product Characteristics TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP.TMB-D Blotting liquid ready-to-use substrate is a highly active and stable blotting substrate utilized for measuring HRP probe activity. A stable blue precipitate is formed at the reaction site. The substrate does not contain NMP (1-methyl-2- pyrrolidone) making it REACH Restricted Substances List Annex XVII compliant, while ensuring maximal safety during use, and minimal waste problems after use.Composition & Properties Ready-to-use substrate: Includes substrate buffer and hydrogen peroxide. No other reagents should be added.Working Procedure The following procedure is applicable to nitrocellulose membranes. The procedure must be optimized for other membranes.1.The desired amount of substrate is poured into a sealed container and allowed to reach room temperature, in the dark, before use. 2.After the last incubation with HRP-labelled Streptavidin or HRP-labelled secondary antibody it is recommended to wash the membrane in a 0.1 M Tris buffer pH 7.4.3.Shake off the excess buffer and incubate the membrane in the TMB-D Blotting solution for 10 minutes. 4.Wash the membrane in distilled water and allow it to dry. 5.The site of positive reaction will appear light blue with no or very little background staining.Tips & Tricks • The membrane can be blocked with Kementec’s Synthetic Blocking Buffer for Blotting, (cat. no. S494457). • For long-term preservation of the results, the membranes must be stored in the dark.Handling & Storage • Store solution at 2-8⁰C in the dark. • Avoid exposure to light, heat and contamination with metal ions or peroxidase. • Re-dispense only into bottles made of High-Density Polyethylene (HDPE), amber color. Dispensing guidelines are available upon request... Read More |