4-Methylumbelliferyl-β-D-Galactoside (Synonyms: 4-Methylumbelliferyl-β-D-Galactopyranoside) |
| Catalog No.GC49114 |
4-Methylumbelliferyl-β-D-Galactoside is a fluorescent substrate that is used for the enzyme release assay to detect human NK cell activity.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 6160-78-7
Sample solution is provided at 25 µL, 10mM.
4-Methylumbelliferyl-β-D-Galactoside is a fluorescent substrate that is used for the enzyme release assay to detect human NK cell activity[1]. 4-Methylumbelliferyl-β-D-Galactoside is hydrolyzed during the enzymatic reaction, and the released 4-methylumbelliferone could be quantified by fluorometry [2]. 4-Methylumbelliferyl-β-D-Galactoside can act as the substrate for rapid detection of total and fecal coliform bacteria (TC and FC) in drinking water [3]. 4-Methylumbelliferyl-β-D-Galactoside is currently used to develop a series of highly sensitive assays to probe β-D-galactoside activity in cells[4].
References:
[1] Ohmori H, Ikeda H, Tanigawa T, et al. Enzyme release assay of human NK cell activity using β-galactosidase-expressing K562 target cell line[J]. Journal of immunological methods, 1993, 164(1): 131-135.
[2] Hattenberger M, Mascher F, Kalcher K, et al. Improved method for the fluorimetric detection of β-D-galactosidase in water[J]. International journal of hygiene and environmental health, 2001, 203(3): 281-287.
[3] Berg J D, Fiksdal L. Rapid detection of total and fecal coliforms in water by enzymatic hydrolysis of 4-methylumbelliferone-beta-D-galactoside[J]. Applied and environmental microbiology, 1988, 54(8): 2118-2122.
[4] Ramsay J P. High-throughput β-galactosidase and β-glucuronidase assays using fluorogenic substrates[J]. Bio-protocol, 2013, 3(14): e827-e827.
Experimental protocol for β-galactosidase activity detection based on 4-Methylumbelliferyl-β-D-Galactoside
This experimental protocol is derived from the literature. Please make modifications according to your specific requirements.
1.Solution preparation
(1) Substrate solution:
2.5mg of 4-methylumbelliferyl-β-D-galactoside was weighed and dissolved in 1ml of citrated acid-sodium phosphate buffer (pH 4.5), stored in the dark at 4°C, and prepared immediately for use.
(2) 4-methylumbelliferone (4-MU) reference material Solution series:
Stock solution A (1mM): 4-MU 1.98mg dissolved in 10ml distilled water and diluted to different concentrations: 0, 0.1, 0.5, 1, 5, 10μM.
(3) Reaction buffer:
Citrate-sodium phosphate buffer (50mM, pH 4.5) containing 0.45% NaCl.
2. Sample preparation
Jurkat cells were harvested, washed twice with PBS, added cell lysate, lysed at 4°C for 30min, centrifuged at 12,000rpm for 10min, and the supernatant was removed for protein quantification.
3. Enzyme Assay:
The enzymatic reaction was initiated by mixing:
(1) 250μl lysed cell suspension with 0.1% albumin (220μl cell suspension in PBS lysed immediately after adding 30μl cell lysis buffer).
(2) 750μl 4-Methylumbelliferyl-β-D-Galactoside in 50mM citrate-phosphate buffer, pH 4.5 with 0.45% NaCl. The final substrate concentration in the reaction mixture was 0.8mM.
(3) The mixture was incubated at 37°C for 30min. After the time, the enzymatic reaction was terminated by adding 10ml 100mM glycine-NaOH buffer, pH 10.7. The same protocol was used for the blank mixture.
(4) The relative fluorescence (excitation 365nm, emission 445nm) was measured using a spectrofluorimeter. The results were compared with a 4-MU standard curve. The enzymatic activity was expressed as μU per number of cells, where U is the amount of enzyme that catalyzes the conversion of 1μM of substrate per 1min.
References:
[1] Kwapiszewski R, Szczudlowska J, Kwapiszewska K, et al. Determination of acid ?-galactosidase activity: methodology and perspectives[J]. Indian Journal of Clinical Biochemistry, 2014, 29(1): 57-62.
| Cas No. | 6160-78-7 | SDF | |
| Synonyms | 4-Methylumbelliferyl-β-D-Galactopyranoside | ||
| Canonical SMILES | CC(C1=CC=C(O[C@@H]2O[C@@H]([C@@H]([C@@H]([C@H]2O)O)O)CO)C=C1O3)=CC3=O | ||
| Formula | C16H18O8 | M.Wt | 338.3 |
| Solubility | 50 mg/ml in DMSO (Need ultrasonic), Ethanol: slightly soluble, Methanol: slightly soluble | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 2.956 mL | 14.7798 mL | 29.5596 mL |
| 5 mM | 591.2 μL | 2.956 mL | 5.9119 mL |
| 10 mM | 295.6 μL | 1.478 mL | 2.956 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 27 reference(s) in Google Scholar.)
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