各种缓冲液.pdfVIP

STOCK SOLUTION RECIPIES: Tris-HCl Buffer 10X Tris-HCl (0.5M Tris Base, pH7.6): Trizma Base 61 g Distilled water 1000 ml Adjust pH 7.6 using concentrated HCl Store this solution at room temperature. Dilute 1:10 with distilled water before use and adjust pH if necessary. 20X Tris-HCl (1M Tris Base, pH7.6): Trizma Base 122 g Distilled water 1000 ml Adjust pH 7.6 using concentrated HCl Store this solution at room temperature. Dilute 1:20 with distilled water before use and adjust pH if necessary. 10X Tris-HCl-Tween 20 (0.5M Tris Base, 0.5% Tween 20, pH7.6): Trizma Base 61 g Distilled water 1000 ml Adjust pH 7.6 using concentrated HCl and then add 5 ml of Tween 20. Store this solution at room temperature. Dilute 1:10 with distilled water before use and adjust pH if necessary. Note: Tris-HCl Buffer is used for specific cases of immunohistochemical staining. *** OR you can use Tris Base to make Tris-HCl (note that Tris base is different from Trizma) Tris is a chemical with basic properties, having a pKa of 8.1. It can be used to buffer solutions from drastic pH changes, keeping them in the pH range of 7.0 to 9.0. Make any Tris-HCl buffer in this pH range, at any molarity using these simple steps 1) Calculate Moles of Tris Base mol/L * L = moles needed 2) Calculate Mass of Tris Base Determine the mass of Tris base to weigh by multiplying the number of moles by the molecular weight (121.14 g/mol) of Tris. moles needed * g/mol = g 3) Dissolve Tris Base in Water Dissolve the required mass of Tris into a volume of deionized water approximately 1/3 of the desired volume of buffer to be made. 4) Adjust the pH Using a pH meter, titrate the solution of Tris with 1M hydrochloric acid (HCl) until the correct pH is reached. 5) Bring to Volume Add the TrisHCl mixture to a volumetric flask