Tris-buffered saline

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Tris-buffered saline (TBS) is a buffer used in some biochemical techniques to maintain the pH within a relatively narrow range. Tris (with HCl) has a slightly alkaline buffering capacity in the 7–9.2 range. The conjugate acid of Tris has a pKa of 8.07 at 25 °C. The pKa declines approximately 0.03 units per degree Celsius rise in temperature. This can lead to relatively dramatic pH shifts when there are shifts in solution temperature. Sodium chloride concentration may vary from 100 to 200 mM, tris concentration from 5 to 100 mM and pH from 7.2 to 8.0. A common formulation of TBS is 150 mM NaCl, 50 mM Tris-HCl, pH 7.6. TBS can also be prepared by using commercially made TBS buffer tablets or pouches. [1]

Applications

TBS is isotonic and non-toxic. It can be used to dilute substances used in laboratory experiments. Additives can be used to add to a compound's functionality.

TBS is often used in immuno-blotting for both membrane washing and antibody dilution. [2]

Related Research Articles

Acid Type of chemical substance

An acid is a molecule or ion capable of either donating a proton (i.e., hydrogen ion, H+), known as a Brønsted–Lowry acid, or, capable of forming a covalent bond with an electron pair, known as a Lewis acid.

Agarose

Agarose is a polysaccharide, generally extracted from certain red seaweed. It is a linear polymer made up of the repeating unit of agarobiose, which is a disaccharide made up of D-galactose and 3,6-anhydro-L-galactopyranose. Agarose is one of the two principal components of agar, and is purified from agar by removing agar's other component, agaropectin.

pH Measure of the acidity or basicity of an aqueous solution

In chemistry, pH (, historically denoting "potential of hydrogen" (or "power of hydrogen") is a scale used to specify the acidity or basicity of an aqueous solution. Acidic solutions (solutions with higher concentrations of H+ ions) are measured to have lower pH values than basic or alkaline solutions.

Calcium carbonate Chemical compound

Calcium carbonate is a chemical compound with the formula CaCO3. It is a common substance found in rocks as the minerals calcite and aragonite (most notably as limestone, which is a type of sedimentary rock consisting mainly of calcite) and is the main component of eggshells, snail shells, seashells and pearls. Calcium carbonate is the active ingredient in agricultural lime and is created when calcium ions in hard water react with carbonate ions to create limescale. It has medical use as a calcium supplement or as an antacid, but excessive consumption can be hazardous and cause hypercalcemia and digestive issues.

A buffer solution is an aqueous solution consisting of a mixture of a weak acid and its conjugate base, or vice versa. Its pH changes very little when a small amount of strong acid or base is added to it. Buffer solutions are used as a means of keeping pH at a nearly constant value in a wide variety of chemical applications. In nature, there are many systems that use buffering for pH regulation. For example, the bicarbonate buffering system is used to regulate the pH of blood, and bicarbonate also acts as a buffer in the ocean.

An acid dissociation constant, Ka, is a quantitative measure of the strength of an acid in solution. It is the equilibrium constant for a chemical reaction

Tris Chemical compound

Tris, or tris(hydroxymethyl)aminomethane, or known during medical use as tromethamine or THAM, is an organic compound with the formula (HOCH2)3CNH2. It is extensively used in biochemistry and molecular biology as a component of buffer solutions such as in TAE and TBE buffers, especially for solutions of nucleic acids. It contains a primary amine and thus undergoes the reactions associated with typical amines, e.g. condensations with aldehydes. Tris also complexes with metal ions in solution. In medicine, tromethamine is occasionally used as a drug, given in intensive care for its properties as a buffer for the treatment of severe metabolic acidosis in specific circumstances. Some medications are formulated as the "tromethamine salt" including hemabate (carboprost as trometamol salt), and "ketorolac trometamol".

A lysis buffer is a buffer solution used for the purpose of breaking open cells for use in molecular biology experiments that analyze the labile macromolecules of the cells. Most lysis buffers contain buffering salts and ionic salts to regulate the pH and osmolarity of the lysate. Sometimes detergents are added to break up membrane structures. For lysis buffers targeted at protein extraction, protease inhibitors are often included, and in difficult cases may be almost required. Lysis buffers can be used on both animal and plant tissue cells.

Saline (medicine) Saline water for medical purposes

Saline is a mixture of sodium chloride (salt) and water and has a number of uses in medicine. Applied to the affected area it is used to clean wounds, help remove contact lenses, and help with dry eyes. By injection into a vein it is used to treat dehydration such as from gastroenteritis and diabetic ketoacidosis.

TBE or Tris/Borate/EDTA, is a buffer solution containing a mixture of Tris base, boric acid and EDTA.

Phosphate-buffered saline is a buffer solution commonly used in biological research. It is a water-based salt solution containing disodium hydrogen phosphate, sodium chloride and, in some formulations, potassium chloride and potassium dihydrogen phosphate. The buffer helps to maintain a constant pH. The osmolarity and ion concentrations of the solutions match those of the human body (isotonic).

TE buffer is a commonly used buffer solution in molecular biology, especially in procedures involving DNA, cDNA or RNA. "TE" is derived from its components: Tris, a common pH buffer, and EDTA, a molecule that chelates cations like Mg2+. The purpose of TE buffer is to solubilize DNA or RNA, while protecting it from degradation.

Tricine Chemical compound

Tricine is an organic compound that is used in buffer solutions. The name tricine comes from tris and glycine, from which it was derived. It is a white crystalline powder that is moderately soluble in water. It is a zwitterionic amino acid that has a pKa1 value of 2.3 at 25 °C, while its pKa2 at 20 °C is 8.15. Its useful buffering range of pH is 7.4-8.8. Along with bicine, it is one of Good's buffering agents. Good first prepared tricine to buffer chloroplast reactions.

MES (buffer) Chemical compound

MES is the common name for the compound 2-(N-morpholino)ethanesulfonic acid. Its chemical structure contains a morpholine ring. It has a molecular weight of 195.2 and the chemical formula is C6H13NO4S. Synonyms include: 2-morpholinoethanesulfonic acid; 2-(4-morpholino)ethanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; 2-(4-morpholino)ethanesulfonic acid; MES; MES hydrate; and morpholine-4-ethanesulfonic acid hydrate. MOPS is a similar pH buffering compound which contains a propanesulfonic moiety instead of an ethanesulfonic one.

Radioimmunoprecipitation assay buffer is a lysis buffer used to lyse cells and tissue for the radio immunoprecipitation assay (RIPA). This buffer is more denaturing than NP-40 or Triton X-100 because it contains the ionic detergents SDS and sodium deoxycholate as active constituents and is particularly useful for disruption of nuclear membranes in the preparation of nuclear extracts. The RIPA buffer gives low background but can denature kinases.

In molecular biology, TBST is a mixture of tris-buffered saline (TBS) and Polysorbate 20. It is a buffer used for washing nitrocellulose membrane in western blotting and microtiter plate wells in ELISA assays. Tris is a pH buffer that maintains a pH that usually lies around pH=8. Buffered Saline is a salt that provides an isotone solution. Tween is a soap that removes fat droplets from the blot or well.

Borate buffered saline is a buffer used in some biochemical techniques to maintain the pH within a relatively narrow range. Borate buffers have an alkaline buffering capacity in the 8–10 range. Boric acid has a pKa of 9.14 at 25°C.

A balanced salt solution (BSS) is a solution made to a physiological pH and isotonic salt concentration. Solutions most commonly include sodium, potassium, calcium, magnesium, and chloride. Balanced salt solutions are used for washing tissues and cells and are usually combined with other agents to treat the tissues and cells. They provide the cells with water and inorganic ions, while maintaining a physiological pH and osmotic pressure.

TAPS (buffer) Chemical compound

TAPS is a chemical compound commonly used to make buffer solutions.

SDS-PAGE Biochemical technique

SDS-PAGE is a discontinuous electrophoretic system developed by Ulrich K. Laemmli which is commonly used as a method to separate proteins with molecular masses between 5 and 250 kDa. The combined use of sodium dodecyl sulfate and polyacrylamide gel allows to eliminate the influence of structure and charge, and proteins are separated solely on the basis of differences in their molecular weight.

References

  1. Medicago AB, (2010) Tris buffered saline specification sheet
  2. Salameh, Samer; Nouel, Dominique; Flores, Cecilia; Hoops, Daniel (2018). "An optimized immunohistochemistry protocol for detecting the guidance cue Netrin-1 in neural tissue". MethodsX. 5: 1–7. doi:10.1016/j.mex.2017.12.001. PMC   5737949 . PMID   29276694.