Tris, Tris- HCl Buffer use summary

Tris(hydroxymethyl)aminomethane, Trizma or Tris is a​​​​ zwitterionic buffer which is mainly used as a biological buffer in Neutral or Basic PH. It is extensively used in biochemistry and molecular biology. Other uses are protein electrophoresis, western blotting, anion exchange chromatography, evaluation of bacterial endotoxins^, capillary electrochromatography and Immunoassay test medias.

The PH of a Tris buffer increases when it being cooled. For example, 0.5 M Tris base buffer has PH 6.9 at 37 C° while its PH at 4C° is 7.7. Tris can inhibit some enzymes activity with metal chelating, so should be used with care when studying enzymes. In Immunoassay tests it increases Recovery test for most analytes and may result in ELISA high signal problems. In combination with detergents this issue is worsening. but still, it is proper for ELISA Sample diluent, ELISA Conjugate Buffer, Standards and wash buffer.

Tris Buffer is a very effective bufferic system in combination with EDTA and EGTA in molecular biology. It is common in Lysis buffers, DNA reaction buffer and electrophoresis. low cost, make Tris and Tris-HCL one of the most common buffers in the biology and biochemistry laboratory.

Protocol for 0.1 M Tris Base Buffer PH=7.4

For a 0.1 M solution of Tris Base Buffer, dissolve 12.1 g of Tris base in 950 mL of distilled water.

1. 1. 2. Adjust the pH to 7.4 value by slowly adding approximately 6-7 mL concentrated HCl.
      3. Adjust the volume of the solution to 1000 mL with distilled water.
      4. If necessary, Filter Tris buffer with 0.22 μm sterile filter.

Note1:  Adding concentrated HCl to the Tris buffer can increase the temperature of the solution and change the PH.

Note2: For Immunoassay tests, you do not need to sterilize.

Note3: Tris Base buffer is stable at 4C° at least for 2 weeks.

Protocol for preparation 0.05 M Tris Base Buffer PH=7.4

For a 0.05M solution of Tris Base Buffer, dissolve 6.05 g of Tris base in 950 mL of distilled water.

1. 1. 2. Adjust the pH to 7.4 value by slowly adding approximately 6-7 mL concentrated HCl.
      3. Adjust the volume of the solution to 1000 mL with distilled water.
      4. If necessary, Filter Tris buffer with 0.22 μm sterile filter.

Note1:  Adding concentrated HCl to the Tris buffer can increase the temperature of the solution and change the PH.

Note2: For Immunoassay tests, you do not need to sterilize.

Note3: Tris Base buffer is stable at 4C° at least for 2 weeks.