Tris, buffer use

In addition to the pH, the osmolarity of the buffer used to dilute the primary antibody tends to influence the immunoreactivity of monoclonal antibodies. The changes in the molarity of Tris buffer used for diluting monoclonal antibodies are expected to result in changes in the immunoreactivity of antibodies. It has been reported that the higher the concentration of cations (e.g., Na+) in the buffer or the higher the pH in their presence, the less the immunoreactivity of the monoclonal antibodies (Boenisch, 1999). However, polyclonal antibodies may not show such an adverse effect. 

According to another paper, methamidophos and phosphorus-containing insecticides were detected by beef liver esterases in Tris buffer using indoxyl acetate, K3pe(CN)6, and K4Fe(CN)g. The insecticides gave white spots on a blue background with a detection limit of 15 ng/spot (162). 

Fig. 8. The effect of addition of TPNH after tyrosine formation was stopped. Tris buffer used in place of phosphate buffer.

DEAE-cellulose chromatography. The 50% ethanol solution is poured onto a column of DEAE cellulose (2.6 x 10 cm), and F adsorbed at the top is eluted with 20 mM Tris buffer, pH 7.5, containing 0.2 M NaCl. A low pressure of argon gas is applied to accelerate the flow rate. The fractions containing F are combined, concentrated, and desalted using ethanol. 

The solution is dialyzed against the same buffer using a hollow fiber assembly, and then added onto a column of Affi-Gel Blue (50-100 mesh, 2 x 15 cm, Bio-Rad) prepared with the same buffer. The column is washed with the same buffer. Then luciferase is eluted with 50 mM Tris-HCl, pH 8.5, containing 5mM EDTA, 3 mM DTT, and 0.5 M NaCl (Hastings and Dunlap, 1986, state that it may be preferable to omit the Affi-Gel step because of difficulties encountered). 

C18-0134. One of the most common buffers used in protein chemistry is a weak base called TRIS (the nitrogen atom of the amino group is the basic portion of the molecule) ... 

The kinetics were followed by measuring the increase in absorbance at 400 nm due to the formation of the p-nitrophenoxide anion in a tris buffer solution at pH 8.5. The substrate was used in excess over the free base catalyst, whose concentration was calculated from spectrophotometric data. 

Saito et al.17 Aldehyde-fixed Using the cultured AR in Tris buffer... 

If this antibody was successfully used, to repeat the protocol of AR-IHC with certain tested tissue slides may be necessary to demonstrate the IHC result. If IHC staining result is not satisfactory, further steps are followed by testing various AR solutions with different pH. For convenience, try to use commonly used solutions such as citrate buffer of pH 6.0,Tris-HCl buffer plus EDTA at pH 8 or 9,0.05% citraconic anhydride at pH 7.4. In some cases, low pH of 1-2 (acetic buffer or other type of buffer solutions) may be tested in order to find out the optimal pH value that produces the best IHC result. 

Dissolve the sulfhydryl-containing protein or macromolecule to be modified at a concentration of l-10mg/ml in 50mM Tris, 0.15M NaCl, 5mM EDTA, pH 8.5. EDTA is present to prevent metal-catalyzed oxidation of sulfhydryl groups. The presence of Tris, an amine-containing buffer, should not affect the efficiency of sulfhydryl modification. Not only do amines generally react slower than sulfhydryls, the amine in Tris buffer is of particularly low reactivity. If Tris does pose a problem, however, use 0.1M sodium phosphate, 0.15M NaCl, 5mM EDTA, pH 8.0. 

Add ethanolamine to the particle suspension at a final concentration of 0.1 M to quench any remaining active groups and react with mixing for 1 hour. Other amine-containing quenchers may be used, too, such as Tris buffer. Note DSC-activated sites on the particles that completely hydrolyze will revert back to the original hydroxyls. 

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