CAS 5625-37-6 Good Buffer Solutions PIPES Piperazine-1,4-Diethanesulfonic Acid

Good Buffer Solutions PIPES Piperazine-1,4-Diethanesulfonic Acid CAS5625-37-6 White crystal powder Purity>99%

Keywords: Good’s buffer, PIPES, piperazine-1,4-diethanesulfonic acid

The full name of PIPES is piperazine-1,4-diethanesulfonic acid, a substance that can maintain the pH of the solution within a certain range. When a certain amount of acid and alkali is added to some solutions, it has the effect of hindering the pH change of the solution, which is called a buffering biological buffer. Such a solution is called a buffer solution. PIPES is an important member of Good’s buffer. Good’s buffer is a buffer system dedicated to life science research, which does not participate in and does not interfere with biochemical processes.

Basic information

Molecular Structure

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Application of PIPES

It can not form stable complexes with most metal ions and is suitable for buffer in solution system containing metal ions. It was applied to the purification of tubulin by using cellulose phosphate chromatography. The purified recombinant GTP binding protein ARF1 and ARF2 were purified by gel filtration and used as buffer to crystallize ketoenzyme from E. coli. In addition, because PIPES can form free radicals, it is not suitable for redox system. Low concentration of PIPES buffer solution should be used in cation exchange chromatography because PIPES has relatively high ionic strength and its pKa value is concentration dependent.

Advantage of PIPES

The purity (> 99%) is water-soluble, the process is stable, and the appearance of the product can be guaranteed to be pure white crystal powder.

Piperazine diethanesulfonic acid PIPES has a variety of buffer systems, among which the buffer solution Transformation Buffer (also called TB solution) can be used to prepare highly efficient competent cells.

Competent state refers to a special physiological state in which cells (or bacteria) can take up DNA molecules from the surrounding environment and are not easily broken down by restriction endonucleases in the cell. Some bacteria are naturally competent, such as Bacillus subtilis, and some are artificially competent, such as Escherichia coli. Treating the recipient cells with a special method can change the permeability of the cell membrane, making it a competent cell that allows carrier molecules with foreign DNA to pass through.

Competent TB solution raw material-PIPES powder

Transformation Buffer preparation:

The transformation of competent bacterial cells uses the calcium chloride method. E. coli taken out in liquid nitrogen or low-temperature refrigerator, streak on LB plate, 37 degrees overnight until a single colony grows. Pick a colony with a diameter of 1-3mm to inoculate the SOB medium in a 250ml/3L Erlenmeyer flask or 80ml/1L Erlenmeyer flask. Incubate at 18 degrees at 150-250RPM for 19-50h. Stop the culture when the OD600 is about 0.4-0.8, cool in 4 degrees ice water for 10 minutes, centrifuge at 300 RPM for 15 minutes, recover the bacteria after removing the supernatant, suspend in 1/3 volume of ice-cold TB solution, and centrifuge again for 10 minutes to recover the bacteria. Suspend with 1/12.5 volume of TB, add DMSO with a final concentration of 7%, and then cool for 10 minutes in 0.1-1ml aliquots, and store directly in liquid nitrogen or -80°C.

In the TB solution of suspended Escherichia coli, the main components include buffer PIPES and calcium chloride to transform cells into competent. It should be noted that this method is suitable for E. coli strains that do not contain plasmids, and is not suitable for large plasmids. The transformation efficiency may be reduced after cryopreservation. In addition, the optimum OD600 of different bacterial strains is also different, and needs to be adjusted according to different experiments.

There are many methods for preparing competent cells with TB, and the efficiency of different methods may be different, and the applicable strains or cell types may also be different. The PIPES produced by Desheng is a powder reagent, and competent TB solutions can be prepared according to different needs.