Application and Preparation of CAPS
2019-04-26 09:49:04
CAPS (N-Cyclohexyl-3-Aminopropanesulfonic Acid) is one of the commonly used Good's buffers in separation of basic drugs by enzymatic chemistry and HPLC. It has a pKa value of 10.4 at 25°C and a pH buffer range of 9.7~11.1. What specific experiments it can be used? How to prepare it?
It can be used in the following experiments:
(1) Western blot transfer buffer or transmembrane buffer;
(2) Binding buffer and eluent in cation exchange chromatography;
(3) Running buffer in capillary electrophoresis;
(4) Effective crystallization solution for various proteins;
(5) Enzyme assay buffer;
(6) Supporting alkaline phosphatase activity and inhibit Aeromonas growth at pH 10.5;
(7) The determination of bicinchoninic acid (BCA).
The preparation method (CN106117088A):
(1) Preparation of catalyst cerium light rare earth doped nano titanium dioxide: titanium powder and cerium light rare earth are mixed at a weight ratio of 10:1, calcined in a muffle furnace at a temperature of 650°C for 15h, and then dissolved in distilled water. It was shaken in a water bath, mixed by shaking at 65°C for 60 min, and finally filtered and dried.
(2) The sulfite, bisulfite and propenol are reacted under the special catalyst to prepare 1,3-propanesulfonate, the reaction temperature is 65~90°C, the pH is 6~6.5, and the reaction time is 4~12 hours, the reaction molar concentration ratio is: propenol:bisulfite:sulfite = 1:1.1~1.7:0.1~0.6;
(3) The obtained 1,3-propanesulfonate reacted with carbonic acid at a temperature of 30~50°C for 1~2 h to obtain 1,3-propanesulfonic acid;
(4) 1,3-propanesulfonic acid and cyclohexylamine are reacted in a water bath at a temperature of 70~90°C for 2~5 h to obtain a crude product of 3-cyclohexylamine-1-propanesulfonic acid;
(5) The crude product is dehydrated by molecular distillation technique at a temperature of 140~150°C and a pressure of 13~266 Pa;
(6) Washing with acetone for 3~7 times to remove cyclohexylamine, and finally washing 3-cyclohexylamine with distilled water for 3~7 times, and 500 mL of distilled water was used each time to obtain purified 3-cyclohexylamine-1-propanesulfonic acid.
Most protein experiments should be performed at 4°C to prevent protein degradation, the configured buffer needs to be sealed and stored, used up as soon as possible after opening.
Edited by Suzhou Yacoo Science Co., Ltd.