Reaction equation
[CH3:1][CH:2](C[C@H](NC([C@@H](NC=O)CCSC)=O)C(N[C@H](C(O)=O)CC1C=CC=CC=1)=O)[CH3:3].CO[C:33]1C=C(C2C=CC(N)=C(OC)C=2)C=C[C:34]=1N.[OH:49]O.[C:51]([O-:63])(=O)[CH2:52][C:53]([CH2:58][C:59]([O-])=O)([C:55]([O-:57])=O)O>>[CH3:33][C:34]1[C:51](=[O:63])[C:52]2[CH:1]=[CH:2][CH:3]=[C:55]([OH:57])[C:53]=2[C:58](=[O:49])[CH:59]=1Participants
Conditions
- Temperature
- 37 °C
Procedure
Eppendorf tubes containing either 25 mg of CPPD or 1 uM fMLP (with 0.5 uM cytochalasin B) were maintained at 37° C. To the tubes were added 0.5 mL of cells at 37° C. followed by shaking to initiate the reactions. At appropriate times, tubes were centrifuged at 10,000× g for 10 s and 0.4 mL of supernatant was stored at −20° C. for later assay. Myeloperoxidase (MPO) activity was measured by the increase in absorbance at 450 nm that accompanies the oxidation of o-dianisidine. Dianisidine (7.8 mg) was dissolved in 100 mL of 0.1 M citrate buffer, pH 5.5 and to a 1 mL cuvette were added 0.89 mL of the dianisidine solution, followed by 50 mL of 1% Triton X 100, 10 mL of 0.05% hydrogen peroxide and 50 mL of crystal-cell supernatant. MPO activity was determined from the change in absorbance (450 nm) per minute, (ΔA450), using the following relationship: Dianisidine oxidation (nmol/min)=50 ΔA450
Workup
- additionTo the tubes were added 0.5 mL of cells at 37° C.
- customwas stored at −20° C. for later assay
- temperatureincrease in absorbance at 450 nm that
Sources & evidence
- ORD record
- ord-4bb86f3d36404c219fecf74828b3bf26
- Source dataset
- uspto-grants-2014_11
- Patent number
- US08883856B2
Linked notes
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