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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 pK a of 8.07 at 25 °C.
As temperature rises from 25 °C to 37 °C, the pH of a tris buffer will decrease an average of 0.025 units per degree. [9] In general, a 10-fold increase in tris buffer concentration will lead to a 0.05 unit increase in pH and vice versa. [9]
A typical recipe for making 1X TE buffer is: 10 mM Tris, bring to pH 8.0 with HCl; 1 mM EDTA, bring to pH 8.0 with NaOH; TE buffer is also known as T 10 E 1 buffer, which can be read as "T ten E one buffer".
The following is a sample recipe for TBST: 20 mM Tris; 150 mM NaCl; 0.1% Tween 20; Adjust pH with HCl to pH 7.4–7.6 The simplest way to prepare a TBS-Tween solution is to use TBS-T tablets. They are formulated to give a ready to use TBST solution upon dissolution in 500 ml of deionized water. [1]
54 g of Tris base (CAS# 77-86-1, free base) 27.5 g of boric acid (CAS# 10043-35-3) 20 ml of 0.5 M EDTA (CAS# 60-00-4) (pH 8.0) Adjust pH to 8.3 by HCl. [2] TBE can be diluted to 1X prior to use in electrophoresis, 0.5x is acceptable as well. Higher concentrations will result in poor results due to excessive heat generation.
Buffer capacity falls to 33% of the maximum value at pH = pK a ± 1, to 10% at pH = pK a ± 1.5 and to 1% at pH = pK a ± 2. For this reason the most useful range is approximately pK a ± 1. When choosing a buffer for use at a specific pH, it should have a pK a value as close as possible to that pH. [2]