Ethylene Glycol → 7-(Benzo(a)pyren-6-yl)guanine

HRID 1458197

反応式

HRID 1458197 の反応式
[CH:1]1[C:14]2=[C:15]3[C:16]4[C:11]([CH:12]=[CH:13]2)=[C:10]2[CH:17]=[CH:18][CH:19]=[CH:20][C:9]2=[CH:8][C:7]=4[CH:6]=[CH:5][C:4]3=[CH:3][CH:2]=1.C[O:22][C:23]1[N:31]=[C:30]([NH2:32])[N:29]=[C:25]2[N:26]=[CH:27][NH:28][C:24]=12.C(O)CO>CO>[CH:1]1[C:14]2=[C:15]3[C:16]4[C:11]([CH:12]=[CH:13]2)=[C:10]2[CH:17]=[CH:18][CH:19]=[CH:20][C:9]2=[C:8]([N:28]2[C:24]5[C:23](=[O:22])[NH:31][C:30]([NH2:32])=[N:29][C:25]=5[N:26]=[CH:27]2)[C:7]=4[CH:6]=[CH:5][C:4]3=[CH:3][CH:2]=1

参加物

6

反応物 4

HCID 174Ethylene Glycol2 回HCID 2336Benzo[a]pyreneHCID 65275O-(6)-Methylguanine2 回HCID 178435178HCID 178435178、化合物

生成物 1

HCID 1356185177-(Benzo(a)pyren-6-yl)guanine

試薬・触媒・溶媒 1

HCID 887Methanol溶媒

実験手順

While the blocking reaction is progressing, NAb (selective for BP-6-N7Gua) and BP-6-N7Gua or benzo[a]pyrene (BP) or methylguanine (Me-N7Gua) or other structures related to BP-6-N7Gua are combined in separate Eppendorf tubes (1.5 mL polypropylene tubes). MAb is diluted 1:500 to 1:5000 in TBS buffer (0.05M Tris, 0.20M NaCl, pH 7.5) containing 10% (v/v) ethylene glycol as a protein stabilizer, then 500 uL is dispensed to each Eppendorf tube seated in a thermal mixing block set at 45 degrees C. These samples are heated for 10 minutes with vortexing. BP-6-N7Gua or BP or Me-N7Gua or other structures related to BP-6-N7Gua are diluted in methanol (MtOH), then added with vortexing in 10 uL volumes to duplicate samples of heated MAb solution. MtOH, alone, is added as a control to selected Eppendorfs. The reaction mixtures are heated with vortexing for an additional 20 minutes, then the reaction mixtures are cooled by addition of 100 uL of TBS buffer with 10% ethylene glycol. The reaction mixtures are incubated an additional 45 minutes at 37 degrees C.

後処理

  1. customWhile the blocking reaction
  2. additionMAb is diluted 1:500 to 1:5000 in TBS buffer (0.05M Tris, 0.20M NaCl, pH 7.5)
  3. customset at 45 degrees C
  4. temperatureThese samples are heated for 10 minutes with vortexing
  5. additionadded with vortexing in 10 uL volumes
  6. additionMtOH, alone, is added as a control
  7. customThe reaction mixtures
  8. temperatureare heated with vortexing for an additional 20 minutes
  9. customthe reaction mixtures
  10. customThe reaction mixtures

出典と根拠

ORDレコード
ord-6f5cb0fabd254de5bf7ac20c419cf965
出典データセット
uspto-grants-1998_12
特許番号
US05849906

関連ノート

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