Buffers - Ready-to-Use Buffers: Tablets and Powders

BES (N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid) is a useful secondary standard biochemical buffer. Useful pH range for BES is 6.4 to 7.8. It is useful for diagnostic assay manufacturing industry. BES, a sulfonic acid-containing cross-linking agent, induces cross-linking between sulfonated polyimide chains in sulfonated polyimide membranes. Synonyms: N,N-Bis(2-hydroxyethyl)-2-aminoethanesulfonic acid; N,N-Bis(2-hydroxyethyl)taurine.

Bicine is recommended for low temperature biochemical work and for the preparation of stable substrate solution for serum guanase determination. Bicine is used for different applications, ranging from buffers used in enzymatic reactions to electrophoretic buffers. Bicine can not be used for BCA and Lowry protein determination procedures. Working concentrations: 3 mM up to 100 mM.

2-[Bis(2-hydroxyethyl)imino]-2-(hydroxymethyl)-1,3-propandiol. Puffersubstanz: Daboo M. and Bates R. (1970) J. Phys. Chem., 74, 702-5. Bis-Tris is an amino buffer very similar in its chemical structure to Trizma ("Tris": e.g. M6210 > or M6215 >). Bis-Tris is a frequently used important buffer if working with proteins or nucleic acids. Bis-Tris is also used very often zu replace buffers that contain Cacodylic acid (Dimethylarsinic acid).

3-(Cyclohexylamino)-1-propane sulfonic acid

CHAPS is a nondenaturing zwitterionic detergent for membrane biochemistry, isoelectric focusing and two-dimensional electrophoresis. Useful for solubilizing membrane proteins and breaking protein-protein interactions. Its small micellar molecular weight (6150 Da) and high critical micellar concentration (6-10 mM) allow it to be removed from samples by dialysis. It is also suitable for protein solubilisation for isoelectric focusing and two-dimensional electrophoresis. CHAPS is commonly used for non-denaturing (without urea) IEF and has been shown to give excellent resolution of some subcellular preparations and plant proteins. Commonly used concentrations for IEF are 2-4% (v/v).

CHAPS is a nondenaturing zwitterionic detergent for membrane biochemistry, isoelectric focusing and two-dimensional electrophoresis. Useful for solubilizing membrane proteins and breaking protein-protein interactions. Its small micellar molecular weight (6150) and high critical micellar concentration (6-10 mM) allow it to be removed from samples by dialysis. It is also suitable for protein solubilisation for isoelectric focusing and two-dimensional electrophoresis. CHAPS is commonly used for non-denaturing (without urea) IEF and has been shown to give excellent resolution of some subcellular preparations and plant proteins. Commonly used concentrations for IEF are 2-4% (v/v).

3-[(3-Cholamidopropyl)-dimethylammonio]-2-hydroxy-1-propane sulfonate

CHES shows a pKa (25°C) of 9,55 which makes CHES useful as a buffering component in the pH range between 8.6 to 12.0. CHES interferes in the protein quantification according to Lowry. It is used in crystallisation of Phosphotriesterase (50mM) or in the chemical modification of Bacteriorhodopsin (10mM). Synonyms are: 2-(Cyclohexylamino)ethanesulfonic acid; 2-(N-Cyclohexylamino)ethanesulfonic acid. Other buffers are: BES, BICIN, MOPS, CAPS, CHAPS

Hepes (N-2-hydroxyethyl) piperazine-N'-2-ethanesulfonic acid) is used in biochemistry, molecular biology, microbiology and cell culture as buffer substance with optimal buffer range at pH6.8-8.2 (pKa = 7.55 (20°C). Hepes is one of the "goods buffers." Dr. Norman Good et al. descriped in his work in 1966 a selection of "biological buffers" that display similar characteristics. These characteristics include: pKa value between 6.0 and 8.0, high solubility, non toxic, limited effect on biochemical reactions, very low absorbence between 240 nm and 700 nm, enzymatic and hydrolytic stability, minimal changes due to temperature and concentration, limited effects due to ionic or salt composition of the solution, limited interaction with mineral cations, and limited permeability of biological membranes. Goods buffers are: BES, BICINE, CAPS, EPPS, HEPES, MES, MOPS, PIPES, TAPS, TES, TRICINE.

1,3-Diaza-2,4-cyclopentadiene, Glyoxalin

Buffering substance. (Good N.E., Izawa S. (1972) Methods Enzymol. 24, 53-68, Sankar M., Bates R.G. (1978) Anal. Chem. 50, 1922-24). Does not interfere with Folin protein assay. MES partially decomposes when autoclaved in the presence of glucose.

Buffering substance. (Good N.E., Izawa S. (1972) Methods Enzymol. 24, 53-68, Sankar M., Bates R.G. (1978) Anal. Chem. 50, 1922-24). Does not interfere with Folin protein assay. MES partially decomposes when autoclaved in the presence of glucose.

MOPS is used as a buffer substance at pH 6.5 to 7.9. (Goods N.E., Izawa S. (1972) Methods Enzymol., 24, 53, Ferguson W.K. et al. (1980) Anal. Biochem., 104, 300).

Buffering substance in the pH range 6,1-7,5. Min. 99.5% pure. Good N.E. et al. (1966) Biochemistry 5, 467-77, Shipman jr. Ch. (1969) Proc. Soc. Exp. Biol. Med. 130, 305-28.

TAPS (N-[Tris(hydroxymethyl)methyl]-3-aminopropanesulfonic acid) is used in biochemistry, molecular biology, microbiology and cell culture as buffer substance with optimal buffer range at pH7.7-9.1 (pKa = 8.4 (20°C). Hepes is one of the "goods buffers." Dr. Norman Good et al. descriped in his work in 1966 a selection of "biological buffers" that display similar characteristics. These characteristics include: pKa value between 6.0 and 8.0, high solubility, non toxic, limited effect on biochemical reactions, very low absorbence between 240 nm and 700 nm, enzymatic and hydrolytic stability, minimal changes due to temperature and concentration, limited effects due to ionic or salt composition of the solution, limited interaction with mineral cations, and limited permeability of biological membranes. Goods buffers are: BES, BICINE, CAPS, EPPS, HEPES, MES, MOPS, PIPES, TAPS, TES, TRICINE. Reference:Good N.E. et al. (1966) Hydrogen Ion Buffers for Biological Research, Biochemistry 5, 467-77. Shipman jr. Ch. (1969) Proc. Soc. Exp. Biol. Med. 130, 305-10.

Synonyme: Tris(hydroxymethyl)aminomethan, THAM, Tris-(hydroxymethyl)-aminomethan, Tris-Base, Trometamol, Tromethamin, Amino-2-(hydroxymethyl)-propan-1,3-diol.

Tris base of highest quality. Purity: min. 99.9%. Useful for buffers of pH7.2 to 9.0. For use in biochemistry and enzymatic research. Tris is the most commonly used buffer in biological research. One of its most important applications is the use as an electrophoresis buffer (e.g. TBE (M3206 Buffer Grade 10-time concentrated, M3088 MolBio Grade 10-time concentrated, M3405 Buffer Grade 5-time concentrated) and D2033 (ready-to-use powder) or TAE (M3085 10-times concentrated and M3087 50-times concentrated) for polyacrylamide and agarose gel electrophoresis, respectively. Besides, Tris is used as TE buffer (pH8.0) for the storage and dissolving of nucleic acids. EDTA protects the nucleic acids by complexing Me++-ions needed from nucleases for their activity. A 1M Tris solution is made up by dissolving 121g Tris base in 800mL of dionized water, adjusting to the desired pH with conc. HCl and filling up to 1 Liter with deionized water. Tris should not be used at pH values under ~pH 7.2 or above ~pH 9.0. The pH value of a Tris buffer strongly depends on the temperature and the pH changes by 0.1 units when diluted by a factor of 10. Therfore, Tris buffers should be prepared at the temperature where it is used and the pH changes should be taken into consideration when the buffer should be diluted. Tris will inactivate the RNase inhibitor DEPC, which is commonly used to treat equipment and solutions, that get into contact with the RNA during its preparation. Synonym: THAM, Tris-(hydroxymethyl)-aminomethane, Tris-Base, Trometamol, Tromethamine, Amino-2-(hydroxymethyl)-propan-1,3-diol. Application and Literature1. Blocking in western blots by incubation of the nitrocellulose membrane after blotting with TBST (10 mM Tris-HCl, pH8.0, 150mM NaCl, 0.05% Tween® 20) with 5% nonfat dried milk (Takemoto, Y. et al. (1995) EMBO J. 14, 3403-3414) or TBS (100mM Tris-HCl, pH7.5, 0.9% NaCl) with 10% (w/v) nonfat dried milk (Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. & Struhl, K. (eds.) (1995) Current Protocols in Molecular Biology. Supplement 39 Page 10.8.10; Greene Publishing & Wiley-Interscience, New York).2. TBE is one of the most commonly used electrophoresis buffers with the composition for a 10X buffer: 890mM Tris base, 890mM boric acid, 20mM EDTA (pH8.3) (for 1Liter: 108g Tris base, 9.3g EDTA x Na2, 55g boric acid, pH8.3). (Peacock, A.C. & Dingman, C.W. (1967) Biochemistry 6, 1818-1827; Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. & Struhl, K. (eds.) (1995) Current Protocols in Molecular Biology. Supplement 40 Page A.2.5, Greene Publishing & Wiley-Interscience, New York; Ogden, R.C. & Adams, D.A. (1987) Methods Enzymol. 152, 61-87).