반응 #2028917

ord-d11127dce50a41d89a5cb6d853e160cd

반응 방정식

O=C1C=Cc2n/c(=C3\N[C@@H](C(=O)O)CS3)sc2=C1
luciferin
O=S(=O)([O-])CCS.[Na+]
MESNA
Nc1ncnc2c1ncn2[C@@H]1O[C@H](COP(=O)(O)OP(=O)(O)O)[C@@H](O)[C@H]1O
ADP
O=C(O)CN(CCN(CC(=O)O)CC(=O)O)CC(=O)O
EDTA
→
Nc1ncnc2c1ncn2[C@@H]1O[C@H](COP(=O)(O)O)[C@@H](O)[C@H]1O
Adenylate

반응물

O=C1C=Cc2n/c(=C3\N[C@@H](C(=O)O)CS3)sc2=C1
luciferin
O=S(=O)([O-])CCS.[Na+]
MESNA
Nc1ncnc2c1ncn2[C@@H]1O[C@H](COP(=O)(O)OP(=O)(O)O)[C@@H](O)[C@H]1O
ADP
O=C(O)CN(CCN(CC(=O)O)CC(=O)O)CC(=O)O
EDTA

시약

없음

용매

NC(CO)(CO)CO
Tris

반응 조건

상세 조건
See reaction.notes.procedure_details.

후처리

  1. 1
    기타Standard curves for the AK enzyme was prepared
  2. 2
    workup.ADDITIONadded to each well
  3. 3
    기타Three separate standard curves
  4. 4
    기타were prepared for incubation of the assay plate at 30 degrees C
  5. 5
    기타50 degrees C
  6. 6
    기타70 degrees C
  7. 7
    workup.ADDITIONwas added

실험 절차

Standard curves for the AK enzyme was prepared as follows. Serial dilutions of purified Sulfolobus acidocaldarius AK from 10 microgrammes/ml to 1 fg/ml were prepared in 50 mM Tris, 25 mM MESNA, pH 7.3. 100 microlitres of enzyme was added to each well of a microtitre plate and 100 microlitres of 135 micromolar ADP 15 mM MgAc, 1 mM EDTA added to each well. Three separate standard curves were prepared for incubation of the assay plate at 30 degrees C., 50 degrees C. and 70 degrees C. for 20 minutes. Following incubation 30 microlitres of luciferin/luciferase reagent (Biothema) was added and the signal read in a luminometer (Orion, Berthold) and the results shown in FIG. 3.

출처

DOI: 10.6084/m9.figshare.5104873.v1특허: US08389208B2uspto-grants-2013_03