反応 #495239

ord-69ecc6a4f49b4677afae8d6f72ac3cfc

反応方程式

CC(C)CCC[C@@H](C)[C@H]1CC[C@H]2[C@@H]3CC=C4C[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C.N[C@H](C(=O)O)[C@H](O)c1ccc(O)c(O)c1
DOPS cholesterol
[Ca+2].[Cl-].[Cl-]
CaCl2
[Ca+2].[Cl-].[Cl-]
CaCl2
[Ca]
calcium
CC(C)CCC[C@@H](C)[C@H]1CC[C@H]2[C@@H]3CC=C4C[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C
cholesterol
→
N[C@H](C(=O)O)[C@H](O)c1ccc(O)c(O)c1
DOPS

反応物

—
ester
CC(C)CCC[C@@H](C)[C@H]1CC[C@H]2[C@@H]3CC=C4C[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C.N[C@H](C(=O)O)[C@H](O)c1ccc(O)c(O)c1
DOPS cholesterol
[Ca+2].[Cl-].[Cl-]
CaCl2
—
mixture
[Ca+2].[Cl-].[Cl-]
CaCl2
[Ca]
calcium
CC(C)CCC[C@@H](C)[C@H]1CC[C@H]2[C@@H]3CC=C4C[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C
cholesterol

試薬

なし

溶媒

O
water

反応条件

詳細条件
See reaction.notes.procedure_details.

後処理

  1. 1
    workup.ADDITIONCochleates containing OVA
  2. 2
    その他were prepared from unilamellar liposomes
  3. 3
    その他converted to unilamellar liposomes by bath
  4. 4
    その他sonication
  5. 5
    濃縮the molar concentration of DOPS constituting the liposomes
  6. 6
    workup.ADDITIONadditional CaCl2 was added
  7. 7
    workup.ADDITIONthat already added

実験手順

Cochleates containing OVA were prepared from unilamellar liposomes comprising a 9:1 mixture (w/w) of synthetic ester lipid 1,2-Dioleoyl-sn-Glycero-3-[Phospho-L-Serine] sodium salt (DOPS lyophilized powder; obtained from Avanti Polar Lipids Inc.; Alabaster, Ala., USA; MW of 810.04; cat #830035P, Lot #181PS-214) and cholesterol (Sigma Chemical Co.; Cat # C8667), by the standard cochleate method of direct addition of CaCl2 as described (Gould-Fogerite and Mannino, 1993). Briefly, a dried film of 20 mg mixture of the DOPS-cholesterol was hydrated in 1 ml of OVA stock solution (10 mg OVA/ml water) and converted to unilamellar liposomes by bath sonication, by avoiding exposure to air as described (Jin et al., 2000). Ten microliter aliquots of 0.1 M CaCl2 stock solution were added to the liposome suspension with mixing in between each addition, till the total concentration of calcium ions equaled to one half the molar concentration of DOPS constituting the liposomes, and then additional CaCl2 was added to achieve a 3 mM excess over that already added, as described (Gould-Fogerite and Mannino, 1993).

出典

DOI: 10.6084/m9.figshare.5104873.v1特許: US08758763B2uspto-grants-2014_06