4-Methylumbelliferyl-β-D-Galactoside (Synonyms: 4-Methylumbelliferyl-β-D-Galactopyranoside) |
Catalog No.GC49114
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4-Methylumbelliferyl-β-D-Galactoside는 인간 NK 세포 활성을 검출하기 위한 효소 방출 분석에 사용되는 형광 기질이다.
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-Galactoside는 인간 NK 세포 활성을 검출하기 위한 효소 방출 분석에 사용되는 형광 기질이다. 효소 반응 중 이 기질이 가수분해되고 방출된 4-methylumbelliferone은 형광 측정법으로 정량할 수 있다[2]. 게다가 음용수 내 총대장균군(TC) 및 분변대장균군(FC)의 신속한 검출을 위한 기질로도 활용된다[3]. 4-Methylumbelliferyl-β-D-Galactoside는 세포 내 β-D-galactosidase 활성을 탐지하기 위한 고감도 분석법 개발에 일반적으로 사용되고 있다[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.
4-Methylumbelliferyl-β-D-Galactoside를 이용한 β-galactosidase 활성 검출 실험 프로토콜 이 실험 프로토콜은 문헌에서 발췌한 것이고 필요에 따라 수정해서 사용하시기 바랍니다.
1.용액 준비
(1) 기질 용액:
4-methylumbelliferyl-β-D-galactoside 2.5 mg을 정확히 저울에 달아 1 ml 시트르산-인산나트륨 완충액(pH 4.5)에 녹이고 차광 후 4°C에 보관하고 사용 직전에 조제합니다.
(2) 4-methylumbelliferone (4-MU) 표준 용액 시리즈:
원액 A (1 mM): 4-MU 1.98 mg을 증류수 10 ml에 녹인 후 다음 농도로 희석합니다: 0, 0.1, 0.5, 1, 5, 10 μM.
(3)반응 완충제:
0.45% NaCl을 포함하는 Citrate-sodium phosphate buffer (50mM, pH 4.5).
2. 검체 준비
Jurkat 세포를 수득하고 PBS로 2회 세척하고 세포 용해물을 첨가하고 4°C에서 30분간 용해하고 12,000rpm에서 10분간 원심분리하고 단백질 정량을 위해 초산액을 제거하였습니다.
3. 효소 검사:
혼합해서 효소반응이 시작되었습니다:
（1）250 μ l lysed 셀 서스펜션과 0. 알부민 (220 μl에서 셀 정학 PBS lysed 추가 한 후 즉시 30 μl 용해 완충 액 셀)다.
(2) 50 mM 시트르산-인산 완충액(pH 4.5, 0.45 % NaCl)에 녹인 4-Methylumbelliferyl-β-D-Galactoside 750 μl을 첨가해서 기질 최종 농도를 0.8 mM로 조정하였습니다.
(3) 반응 혼합액을 37 °C에서 30 min 배양한 후 10 ml 100 mM glycine-NaOH 완충액(pH 10.7)을 추가해서 효소 반응을 중단하였고 블랭크도 동일한 방식으로 처리하였습니다.
(4) 분광형광계를 이용해서 상대 형광을 측정(여기 365 nm, 발광 445 nm)하고 4-MU 표준 곡선과 비교하였습니다. 효소 활성은 세포 수당 μU로 표현하였고 1 U는 1분 동안 1 μM 기질을 전환시키는 효소 양으로 정의하였습니다.
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 | |||
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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 saline, 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% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 27 reference(s) in Google Scholar.)