Halite (NaCl) → 磺基琥珀酰亚胺-6-[3-(2-吡啶基二硫代)丙酰胺]己酸

HRID 324924笔记

反应方程式

HRID 324924 反应方程式
[CH2:1]1[C:6](=[O:7])[N:5]([O:8][C:9]([CH2:11][CH2:12][CH2:13][CH2:14][CH2:15][NH:16][C:17]([CH2:19][CH2:20][S:21][S:22][C:23]2[N:28]=[CH:27][CH:26]=[CH:25][CH:24]=2)=[O:18])=[O:10])[C:3](=[O:4])[CH:2]1[S:29]([O-:32])(=[O:31])=[O:30].[Na+].P([O-])([O-])([O-])=O.[Na+].[Na+].[Na+].[Na+].[Cl-].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.[N-]=[N+]=[N-].[Na+]>OP([O-])(O)=O.OP([O-])([O-])=O.[Na+].[Na+].[Na+].[Cl-].[Cl-].[K+].[K+].O>[N:28]1[CH:27]=[CH:26][CH:25]=[CH:24][C:23]=1[S:22][S:21][CH2:20][CH2:19][C:17]([NH:16][CH2:15][CH2:14][CH2:13][CH2:12][CH2:11][C:9]([O:8][N:5]1[C:6](=[O:7])[CH2:1][CH:2]([S:29]([OH:32])(=[O:30])=[O:31])[C:3]1=[O:4])=[O:10])=[O:18]

参与物

8

反应物 5

HCID 5234Halite (NaCl)HCID 6049乙二胺四乙酸二钠HCID 24243十二水磷酸三钠HCID 33557叠氮化钠HCID 4588467磺基琥珀酰亚胺-6-[3-(2-吡啶基二硫代)丙酰胺]己酸2 次

产物 1

HCID 4588468磺基琥珀酰亚胺-6-[3-(2-吡啶基二硫代)丙酰胺]己酸

试剂、催化剂与溶剂 2

HCID 962聚[6-[4-(4-氰基苯基)苯氧基]己基甲基丙烯酸酯]溶剂HCID 24978514Phosphate Buffered Saline溶剂3 次

实验过程

A 10 mM stock solution of the sulfosuccinimidyl 6-[3′-(2-pyridyldithio)-propionamido]hexanoate (Sulfo-LC-SPDP) is prepared by dissolving 1.3 mg Sulfo-LC-SPDP into 2.07 ml de-ionized water. The conjugation reaction is carried out in phosphate buffered saline (PBS) containing 20 mM sodium phosphate buffer, 150 mM NaCl, 1 mM EDTA and 0.02% sodium azide at pH 7.5. One milligram of lyophilized antibody is dissolved in 450 ml PBS, and 50 ml of Sulfo-LC-SPDP stock solution is added to the antibody solution. The mixture is allowed to react at room temperature for 60 minutes. The sample is applied to a 5 ml desalting polyacrylamide column previously equilibrated with 5 bed volumes (25 ml) of PBS. Fractions are eluted using PBS as the elution buffer, and the protein in the fractions is monitored using a COOMASSIE® Protein Assay (Pierce Chemical Co). Typically, 50 ul of the COOMASSIE® reagent is mixed with 50 ul of each fraction in a micro-titer plate. The COOMASSIE® Blue substrate reacts with the protein, producing a blue color, the intensity of which is dependent upon the amount of protein present in the fraction. The fractions which produce the most intense blue color are those containing the majority of the protein eluted. These fractions are pooled together to produce the disulfide form of the final derivatized product. This is typically the form used for contact printing.

后处理

  1. customThe conjugation reaction
  2. customto react at room temperature for 60 minutes
  3. washFractions are eluted
  4. additionTypically, 50 ul of the COOMASSIE® reagent is mixed with 50 ul of each fraction in a micro-titer plate
  5. customThe COOMASSIE® Blue substrate reacts with the protein
  6. customproducing a blue color
  7. customThe fractions which produce the most intense blue color
  8. washeluted

来源与证据

ORD记录
ord-82ac328fa17f4728b2e1fe4cbf5110b9
来源数据集
uspto-grants-2001_04
专利号
US06221579B1

反应笔记

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