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2-Mercaptoethanol → 4-Methylumbelliferone

HRID 645305
Notes
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Reaction equation

HRID 645305 reaction equation
SCCO.C(N(CC([O-])=O)CC(O)=O)CN(CC([O-])=O)CC(O)=O.[Na+].[Na+].CCCCCCCCCCCCOS([O-])(=O)=O.[Na+].CCC(COC(C(N(CC[NH+](C)C)C)=O)(C1C=CC=CC=1)C1C=CC=CC=1)CC.[Cl-].[CH3:75][C:76]1[C:82]2[CH:83]=[CH:84][C:85]([O:87][C@@H]3O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]3O)=[CH:86][C:81]=2[O:80][C:78](=[O:79])[CH:77]=1>C([O-])([O-])=O.[Na+].[Na+]>[CH3:75][C:76]1[C:82]2[CH:83]=[CH:84][C:85]([OH:87])=[CH:86][C:81]=2[O:80][C:78](=[O:79])[CH:77]=1
ParticipantsProcedureWorkupSources & evidenceReaction notes

Participants

7

Reactant 5

HCID 15672-MercaptoethanolHCID 8759EDTA disodium saltHCID 19153X 100HCID 915534-Methylumbelliferyl beta-D-glucuronideHCID 3423265Sodium Lauryl Sulfate

Product 1

HCID 52805674-Methylumbelliferone

Reagents, catalysts & solvents

1
HCID 10340Sodium CarbonateSolvent

Procedure

To evaluate the activity of various promoters, GUS fluorescence was assayed by the method of Jefferson (1987). Maturing seeds at 17 DAF were homogenized in GUS extraction buffer (50 mM NaPO4 [pH 7.0], 10 mM 2-mercaptoethanol, 10 mM Na2-EDTA, 0.1% SDS, 0.1% Triton X-100). After centrifugation, 10 μl of the supernatant was mixed with 90 μl of assay buffer containing 1 mM 4-methylumbelliferyl-β-D-glucuronide (MUG). After incubating for one hour at 37° C., 900 μl of 0.2 M Na2CO3 was added to the mixture to terminate the reaction. Values obtained using a fluorometer were compared with those obtained from serial dilutions of 4-methylumbelliferone (4 MU). The protein amount was determined using a Bio-Rad Protein Assay kit, with serum albumin as the standard. Three seeds were assayed for each transgenic plant.

Workup

  1. additionAfter centrifugation, 10 μl of the supernatant was mixed with 90 μl of assay
  2. customto terminate
  3. customthe reaction
  4. customValues obtained

Sources & evidence

ORD record
ord-3c96df02940a482cbfe96ab3c1301aea
Source dataset
uspto-grants-2010_04
DOI
10.6084/m9.figshare.5104873.v1
Patent number
US07700835B2

Reaction notes

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On this pageParticipantsProcedureWorkupSources & evidenceReaction notes