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Galactosidase

Galactosidases are a class of enzymes that catalyze the hydrolysis of galactosides, including lactose, into monosaccharides.
These enzymes play a vital role in the metabolism of galactose-containing compounds and have diverse applications in food processing, pharmaceutical development, and biotechnology.
Galactosidases can be derived from various sources, including microorganisms, plants, and animals, and their specific characteristics and activities can vary depending on the source and structure.
Researchers studying galactosidases can optimize their work by utilizing cutting-edge AI-driven platforms like PubCompare.ai, which help identify the best protocols and products for their research, thereby reducing time and effort while improving reproducibility and accuracy.

Most cited protocols related to «Galactosidase»

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Publication 2014
Adipocytes Adipogenesis Biological Assay Cells Cloning Vectors Galactosidase interferon regulatory factor 4, human Luciferases Luciferases, Renilla Mus Paragangliomas 3 Plasmids PPARGC1A protein, human Promega Transfection Trypsin UCP1 protein, human
OS were extracted from human milk obtained from the milk banks in San Jose, CA and Austin, TX. The extraction method was the same as in our previous publication.27 (link), 45 (link) Sodium borohydride (98%) and 2,5-dihydroxybenzoic acid (DHB) were purchased from Sigma-Aldrich (St. Louis, MO). Nonporous graphitized carbon cartridges (GCC, 150mg bed weight, 4mL cartridge volume) were bought from Alltech (Deerfield, IL). Standard HMOs were purchased from Dextra Laboratories (Earley Gate, UK). α(1-2)-Fucosidase was from EMD Calbiochem (La Jolla, CA). β(1-3)-Galactosidase was from New England Biolab (Beverly, MA). β(1-4)-Galactosidase was from ProZyme (San Leandro, CA). α(1-3,4)-Fucosidase was from Sigma-Aldrich (St. Louis, MO). All other reagents were of analytical or HPLC grade.
Publication 2010
2,3-dihydroxybenzoic acid austin Carbon Fucosidase Galactosidase High-Performance Liquid Chromatographies Milk, Human sodium borohydride
The OS used in this study are from pooled human milk provided by milk banks in San Jose, CA and Austin, TX. HMOs were isolated from the milk using a previously described procedure involving defatting, chloroform/methanol extraction, ethanol precipitation, and evaporation.6 (link) A sample enrichment step with solid phase extraction (SPE) employing graphitized carbon cartridge (GCC) was used before the analysis. GCC (150 mg bed weight, 4mL volume) were purchased from Alltech (Deerfield, IL). Sodium borohydride (98%) and 2, 5-dihydroxybenzoic acid (DHB) were obtained from Sigma–Aldrich (St. Louis, MO). Standard HMOs were purchased from Dextra Laboratories (Earley Gate, UK). α(1–2)-Fucosidase was obtained from EMD CALBIOCHEM (La Jolla, CA), β(1–3)-galactosidase and α(2–3)-neuraminidase from New England Biolab (Beverly, MA), β(1–4)-galactosidase from ProZyme (San Leandro, CA), and α(1–3,4)-fucosidase from Sigma–Aldrich (St. Louis, MO). The non-selective sialidase was purified and provided by Prof. David Mills from the Department of Viticulture and Enology in UC Davis. All reagents are of analytical or HPLC grade.
Publication 2011
2,3-dihydroxybenzoic acid austin Carbon Chloroform Ethanol Fucosidase Galactosidase High-Performance Liquid Chromatographies Methanol Milk Milk, Human Neuraminidase sodium borohydride Solid Phase Extraction
The TZM-bl neutralization assay was used to quantify NAb breadth as previously described [45] (link). Briefly, 500 infectious pseudovirus particles, as determined by the infectious titer described above, were incubated in duplicates with 2-fold serial dilutions of plasma for 1 hour, beginning with an initial concentration of 1∶100, before 10,000 TZM-bl reporter cells per well were added. Each plasma-virus combination was tested in duplicate and the assay was repeated twice. Infection levels were determined by B-galactosidase activity after 48 hours using a chemiluminescent readout. The IC50, or reciprocal plasma dilution at which 50% of the virus is neutralized, for each plasma-virus pair was calculated using linear interpolation from the neutralization curve. In this assay, a plasma sample was considered to be below the detectable limit of neutralization for a given virus if the lowest dilution (1∶100) did not show >50% neutralization. With this criterion, plasma samples that showed neutralization below the limit of detection were designated an IC50 value of 50, the midpoint between our starting dilution (1∶100) and 0. Each round of assays included HIV-negative plasma and a HIV-positive plasma pool from 30 HIV-1 infected individuals in Kenya between 1998–2000 [45] (link) serving as negative and positive internal controls, respectively. If a run showed neutralization of the negative control virus (SIVmne CL8) greater than the limit of detection, we considered that a failed run and repeated the assay.
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Publication 2012
Biological Assay Cells Galactosidase HIV-1 HIV Seropositivity Infection Plasma Technique, Dilution Virus
The methodologies used in this study are listed in Table 1 and differences between assay protocols in supplementary Table S1 and supplementary Figure S1. Detailed protocols are available at the EUROPRISE website www.europrise.org. The conventional PBMC based assay [24] (link), [25] (link), [26] (link), [27] (link) with readout based on p24 antigen production involves multiple rounds of virus replication, has a moderate reproducibility and sensitivity, is time-consuming and cumbersome to perform but involves the most physiological target cell. An alternative readout can be the measurement of viral RNA, which shortens the time by several days [28] (link), [29] (link). Intracellular (IC) p24 antigen determination in infected PBMC cultures may be run as a single round assay with increased sensitivity, reproducibility and speed but it is not easy to perform [30] (link). The method of measuring ICp24 was also applied to other target cells, like macrophages [31] (link). Plaque reduction assays use either U87.CD4 or GHOST(3) cells engineered to express coreceptors for HIV [32] (link), [33] (link). In U87.CD4 cells the syncytium-inducing capacity of HIV is exploited, while infected GHOST(3) cells turn green due to the activation of the GFP gene linked to the HIV-2 LTR. These assays are single round, highly reproducible, easy to perform, with sensitivity comparable to the PBMC assay, but require a shorter time. The fusion assay is based on fusion of effector cells expressing the native HIV-1 envelope on their surface (PM1 persistently infected with HIV-1) with target cells expressing the appropriate receptors (initially NIH-3T3 mouse fibroblasts or HeLa human epithelial cells stably expressing human CD4, CCR5 and/or CXCR4). The readout is measurement of ß-galactosidase activity [34] (link). Pseudovirus (PSV)-based assays exist in a number of variant assay formats using different target cells [35] , [36] (link), [37] (link), [38] (link). A selected molecular clone is tested in a single round assay with luciferase readout that results in short-term assays with high reproducibility and sensitivity. Plasmid production and producer cell line culture history are crucial criteria and influences the results. Due to this a fairly large inter-laboratory variation has been documented [23] . Finally, assays using recombinant viruses have also been included [39] (link), [40] (link), [41] (link). This assay type was run with two different starting materials, env sequences were amplified either from culture supernatants or from cloned plasmid.
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Publication 2009
Antigens Biological Assay CCR5 protein, human CD4 Positive T Lymphocytes Cell Lines Cells CXCR4 protein, human Dental Plaque Epithelial Cells Fibroblasts Fusions, Cell Galactosidase Gene Activation Giant Cells HeLa Cells HIV-1 HIV-2 Homo sapiens Hypersensitivity Luciferases Macrophage Mus NIH 3T3 Cells physiology Plasmids Protoplasm Red Cell Ghost RNA, Viral Somatostatin-Secreting Cells Virus Virus Replication

Most recents protocols related to «Galactosidase»

GLA activities were determined in Fabry mouse tissues using previously described methods (Desnick et al., 1973 (link)). In brief, tissue samples were homogenized in chilled reporter lysis buffer (Promega) and protease inhibitor (Pierce) was added to the lysates. Protein concentrations were determined using the Bio-Rad Colorimetric Protein Assay Kit. 10 μL of tissue lysate was added to an equal volume of 10 mM 4-methylumbelliferyl-α-D-galactopyranoside (Sigma-Aldrich), dissolved in assay buffer (0.2 M citrate, 0.4 M phosphate buffer, pH 4.4), and 0.1 M N-acetylgalactosamine (Sigma Aldrich), the latter to inhibit α-galactosidase B activity (Mayes et al., 1981 (link)). Following a 30 min incubation at 37°C, reactions were terminated by the addition of 480 μL of 0.1 M ethylenediamine, pH 10.3. The amount of 4-methylumbelliferone (4-MU) produced was determined by measuring fluorescence using a Synergy H1 fluorometer (BioTek). Tissue α-Gal A activities were expressed as nmol of 4-MU produced per h per mg of total protein (nmol/h/mg). Measurement of plasma GLA activities in wildtype mice for PK studies was performed as described above with the following modifications: lysates were incubated with 5 mM 4-methylumbelliferyl α-D-galactopyranoside in assay buffer [20 mM citrate, 30 mM sodium phosphate (pH 4.4), 0.1 M N-acetylgalactosamine, and 4 mg/mL BSA], and the reaction was stopped by addition of stop buffer (0.1 M Glycine, 0.1 N NaOH], as previously described (Shen et al., 2016 (link)).
AGA activity was measured with 1 mM L-aspartic acid β-(7- amido-4-methylcoumarin) in 10% SuperBlock and 90% 50 mM Tris-HC (pH 7.5) for 60 min at 37°C, and then adding 100 µL of stop buffer [0.2 M glycine, 0.175 M NaOH (pH 10.6)], as previously described (Mononen et al., 1993 (link)). GUSB enzyme assay was performed using 10 mM 4-methylumbelliferyl-β-D-glucuronide (Merck) in 0.1 M sodium acetate (pH 4.6) at 37°C for 30 min, and reactions were stopped by 0.1 M sodium carbonate (Grubb et al., 2008 (link)). GAA activity assay was performed with 3 mM 4-methylumbelliferyl-a-D-glucopyranoside (Merck) in assay buffer (30 mM sodium citrate, 40 mM sodium phosphate dibasic, pH 4.0) at 37°C for 3 h (Flanagan et al., 2009 (link)). Reactions were stopped by the addition of an equal volume of 0.4 M glycine, pH 10.8. IDS activity assay was performed with 2.5 mM 4-Methylumbelliferyl sulfate potassium salt (Merck) in 50 mM sodium acetate, at 37°C for 4 h (Dean et al., 2006 (link)). Reactions were stopped with glycine carbonate buffer (pH 10.7). Fluorescence was measured by microplate reader with 360/40 nm excitation and 440/30 nm emission filters.
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Publication 2023
4-benzaldehydesulfonic acid 4-methylumbelliferyl sulfate, potassium salt 7-methylcoumarin Acetylgalactosamine Aspartic Acid Biological Assay Buffers Carbonates Cardiac Arrest Citrates Colorimetry Enzyme Assays Ethylenediamines Exhaling Fluorescence Galactose Galactosidase Glucuronides Glycine Hymecromone Mice, House Phosphates Plasma Promega Protease Inhibitors Proteins RRAD protein, human Sodium Acetate sodium carbonate Sodium Citrate sodium phosphate Tissues Tromethamine

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Publication 2023
Binding Sites Biological Assay Galactosidase Luciferases Paragangliomas 3 Promega Proteins TATA Box Transfection
To determine the site required for the interaction of MaCFEM85 with MsWAK16, PCR amplification was performed to generate multiple truncated forms of MaCFEM85. The CFEM domain (MaCFEM85-CFEM; aa residues 19–86) and the C terminal without the CFEM domain (MaCFEM85-C, aa residues 87–170) were inserted into the vector pGBKT7 as the bait protein (Supplementary Table S1). Another five variants were constructed using polypeptide synthesis to mutate cysteine to alanine at positions 26, 30, 43, 52, and 26/30/43/50/52/64/69/85 (ΔCFEM8526, ΔCFEM8530, ΔCFEM8543, ΔCFEM8552, and ΔCFEM858 all, respectively). These variants were used to perform Y2H experiments with MsWAK16-ED. The transformed yeast cells were assayed for growth on synthetic dropout SD/-Trp-Leu plates and SD/-Trp-Leu-His-Ade plates containing X-α-Galactosidase (X-α-Gal).
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Publication 2023
5-bromo-4-chloro-3-indolyl beta-galactoside Alanine Anabolism Cells Cloning Vectors Cysteine Galactosidase Polypeptides Proteins Saccharomyces cerevisiae tryptophan-leucine
Prior to the enzymatic assays, the samples were prepared according to the following protocol. Firstly, 0.7 g of sample was suspended in 3.5 mL of 0.2 M phosphate buffer and vortexed thoroughly (Vortex RS-VA 10, Phoenix Instrument, Garbsen, Germany). Secondly, the samples underwent sonification (Time = 2 min, Amplitude = 60, Pulse = 6 s, Cole–Parmer Instrument Co., Vernon Hills, IL, USA). Afterwards, the samples were centrifuged (12,000 rpm, Time = 20 min, Centrifuge MPW-251, MPW, Warszawa, Poland) and the supernatant was transferred to the sterile Eppendorf tubes.
With the use of spectrophotometric methods, the activity of the fecal enzymes α-glucosidase, α-galactosidase, β-glucosidase, β-galactosidase, and β-glucuronidase was determined. The protocols used in the study were based on the reaction of α-glucosidase, β-glucosidase, α-galactosidase, β-galactosidase, and β-glucuronidase with 4-nitrophenyl α-D-glucopyranoside (TCI, Tokyo, Japan), 4-nitrophenyl β-D-glucopyranoside (TCI, Tokyo, Japan), 4-nitrophenyl α-D-galactopyranoside (TCI, Tokyo, Japan), 4-nitrophenyl β-D-galactopyranoside (TCI, Tokyo, Japan), and 4-nitrophenyl β-D-glucuronide (TCI, Tokyo, Japan), respectively.
The used substrates were specific to the respective enzymes present in the fecal sample. The reaction mixture contained 0.5 mL of phosphate buffer (pH = 7, 0.02 M), 0.05 mL of substrate solution (20 mM), and 0.25 mL of sample. Incubation was performed at 37 °C for 15 min (α-glucosidase, α-galactosidase and β-glucuronidase) or 60 min (β-glucosidase and β-galactosidase).
An observed hue shift in the sample to yellow proved the reaction had taken place. The intensity of the color was directly proportional to the amount of p-nitrophenol released. The reactions were inhibited using 0.25 M sodium carbonate after the given reaction time. The absorbance of the samples was measured using the spectrophotometer Rayleigh UV-2601 (BFRL, Beijing, China) at a wavelength of λ = 400 nm. The unit of enzyme activity refers to the amount of p-nitrophenol (expressed in µM) which was released during 1 h of reaction for 1 mg of protein in 1 mL of sample [µMh·mg−1].
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Publication 2023
4-nitrophenyl alpha Glucosidase beta-Galactosidase beta-Glucosidase beta-Glucuronidase Buffers enzyme activity Enzyme Assays Enzymes Feces Galactose Galactosidase Glucuronides Nitrophenols Phosphates Proteins Pulse Rate sodium carbonate Spectrophotometry Sterility, Reproductive
Cell culture, DNA transfection, and cell-based reporter assays were carried out using established protocols [3 (link),55 (link)]. HEK293T cells were selected in this study, as they are easy to transfect and have a high transfection efficiency. HEK293T cells were seeded in a half-area of a 96-well tissue-treated microtiter plate and incubated for 24 h at 37 °C. The cells were transfected with plasmids encoding the TGFBRII receptor gene (100 ng), the SBE-Luc reporter (100 ng), and the pJ7Lac-Z plasmid (50 ng) using GeneJammer transfection reagent (Stratagene, San Diego, CA, USA) following the manufacturer’s instructions. Cells were then treated with drugs at 0.001 µM, 0.01 µM, 1 µM, and 10 µM for 24 h. Twenty-four hours after the treatment, the cells were lysed with 1× Reporter Lysis Buffer (Promega, Madison, WI, USA). Measurement of the luciferase and b-galactosidase was carried out as described elsewhere [3 (link)].
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Publication 2023
Biological Assay Buffers Cell Culture Techniques Cells Galactosidase genejammer Genes Luciferases Pharmaceutical Preparations Plasmids Promega Tissues Transfection

Top products related to «Galactosidase»

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The Senescence-galactosidase staining kit is a laboratory product designed to detect and quantify cellular senescence. It utilizes a histochemical staining method to identify senescent cells by the presence of beta-galactosidase activity at a suboptimal pH.
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The Luciferase Assay System is a laboratory tool designed to measure the activity of the luciferase enzyme. Luciferase is an enzyme that catalyzes a bioluminescent reaction, producing light. The Luciferase Assay System provides the necessary reagents to quantify the level of luciferase activity in samples, enabling researchers to study biological processes and gene expression.
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The Senescence β-Galactosidase Staining Kit is a laboratory tool used to detect and quantify senescent cells. The kit provides reagents and protocols for the histochemical detection of senescence-associated β-galactosidase activity, a widely used biomarker for cellular senescence.
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
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The PGBKT7 is a plasmid vector used for gene expression in yeast cells. It contains a yeast selectable marker and a multiple cloning site for the insertion of DNA sequences.
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β1,4-galactosidase is an enzyme that catalyzes the hydrolysis of β-1,4-linked galactose residues from the non-reducing ends of various oligosaccharides and glycoconjugates. It is commonly used in biochemical and molecular biology applications.
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The PGBKT7 vector is a plasmid used for gene expression in yeast. It contains a GAL1 promoter for inducible gene expression, a TRP1 selectable marker, and a 2-micron origin of replication for high-copy number maintenance in Saccharomyces cerevisiae.
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The PGADT7 is a plasmid vector used in yeast two-hybrid systems for the detection of protein-protein interactions. It contains the GAL4 DNA-binding domain, a multiple cloning site, and various genetic markers for selection in yeast.
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α2-3,6,8 neuraminidase is an enzyme that cleaves α-2,3, α-2,6, and α-2,8 linked sialic acid residues from glycoproteins and glycolipids. It is commonly used in glycobiology research for the removal of sialic acids from carbohydrate structures.
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The Matchmaker Gold Yeast Two-Hybrid System is a laboratory equipment product designed for protein-protein interaction studies. The system utilizes a yeast-based approach to detect and analyze interactions between proteins of interest.

More about "Galactosidase"

Galactosidases, also known as β-galactosidases, are a class of hydrolytic enzymes that play a crucial role in the metabolism of galactose-containing compounds.
These enzymes catalyze the hydrolysis of galactosides, such as lactose, into monosaccharides like glucose and galactose.
Galactosidases are derived from various sources, including microorganisms, plants, and animals, and their specific characteristics and activities can vary depending on the source and structure.
Galactosidases have diverse applications in food processing, pharmaceutical development, and biotechnology.
They are used in the production of dairy products, the treatment of lactose intolerance, and the development of therapeutic enzymes.
Researchers studying galactosidases can optimize their work by utilizing cutting-edge AI-driven platforms like PubCompare.ai, which help identify the best protocols and products for their research, thereby reducing time and effort while improving reproducibility and accracy.
In addition to galactosidases, related enzymes and assays are also important in biological research.
The Senescence-galactosidase staining kit is used to detect senescent cells, while the Luciferase Assay System measures the activity of the luciferase enzyme.
The Senescence β-Galactosidase Staining Kit is another tool used to identify senescent cells, and Lipofectamine 2000 is a transfection reagent used to introduce genetic material into cells.
The PGBKT7 vector and PGADT7 vector are used in yeast two-hybrid systems to study protein-protein interactions, and the β1,4-galactosidase enzyme is involved in the cleavage of galactose-containing glycosides.
The α2-3,6,8 neuraminidase enzyme can be used to remove sialic acid residues from glycoproteins and glycolipids.
By understanding the diverse applications and related technologies associated with galactosidases and related enzymes, researchers can more effectively design and execute their experiments, leading to more robust and reproducible results.