Hello

Tris-HCl can be prepared using Tris base (molecular weight: 121.14 g/mol), or Tris-HCl (Tris base which is already combined with HCl in a 1:1 molar ratio, so the molecular weight is 157.6 g/mol). It is important to note which form you use to make the solution, because the initial pH of the solution you make depends on which form you use.

In this protocol, we will use Tris base (MW 121.14 g/mol) to make 500 mL of 1.0 M Tris-HCl, pH 7.4.

Add 60.57 g of Tris Base to an appropriate beaker (500 mL beaker in this case).

Add 350 mL of deionized water to the beaker.

Add a stir bar to the beaker and leave it on a stir plate until completely dissolved (~1 min).

Begin monitoring pH of the solution. It should be basic (pH >7.4).

Slowly add 32.5 mL of 12 M HCl, while monitoring the pH

Add more HCl as necessary until the pH of the solution reaches 7.4

Once the pH of the solution is 7.4, add enough deionized water to raise the volume to 500 mL.

Sterile-filter, if possible, and store in the refrigerator for up to 4 months.

Citing this Protocol

Farhat, Y. “Reagent Preparation: Tris-HCl (1.0 M, pH 7.4 )"

Good Luck

Hello

Tris-HCl can be prepared using Tris base (molecular weight: 121.14 g/mol), or Tris-HCl (Tris base which is already combined with HCl in a 1:1 molar ratio, so the molecular weight is 157.6 g/mol). It is important to note which form you use to make the solution, because the initial pH of the solution you make depends on which form you use.

In this protocol, we will use Tris base (MW 121.14 g/mol) to make 500 mL of 1.0 M Tris-HCl, pH 7.4.

Add 60.57 g of Tris Base to an appropriate beaker (500 mL beaker in this case).

Add 350 mL of deionized water to the beaker.

Add a stir bar to the beaker and leave it on a stir plate until completely dissolved (~1 min).

Begin monitoring pH of the solution. It should be basic (pH >7.4).

Slowly add 32.5 mL of 12 M HCl, while monitoring the pH

Add more HCl as necessary until the pH of the solution reaches 7.4

Once the pH of the solution is 7.4, add enough deionized water to raise the volume to 500 mL.

Sterile-filter, if possible, and store in the refrigerator for up to 4 months.

Citing this Protocol

Farhat, Y. “Reagent Preparation: Tris-HCl (1.0 M, pH 7.4 )"

Good Luck

Thank you for the quick answer! Is there any reason it should be 1M? I think in our lab they use 0.05M...does molarity make a big difference?

Best regards, Olga

Hi Olga,

Our protocol in our lab as below (little bit same answer by Saleh):

Tris-HCl 1M Solution Ingredient Amount(g)

Trizma base 121.14

HCl (approx) pH7.5 80ml

Make up to 1L with Milli-Q water in a fume hood

All the best,

Ali

This is acommonly used lysis buffer meaning it is a solution used for breaking cells for open in molecular biology experiments that analyse the cellular compounds (e.g. western blot). Most lysis buffers contain salts (e.g. Tris-HCl or EDTA) to regulate the acidity and osmolarity of the lysate.

The molarity depend on how strong/ weak you want this buffer to be. As Saleh and Ali mentioned above that the most commonly used molarity is 1M.

How to Prepare Tris Buffer (25mM= 3.0285 grams Tris in 1 L H2O, pH=7.8)

It is easy to find commercially available tris buffer solution, but it is possible to make it yourself with the appropriate equipment.

Materials:

Calculate the amount of each item you need based on the molar concentration of the solution you want and the quantity of buffer you need.

• tris(hydroxymethyl) aminomethane
• distilled deionized water
• HCl

Procedure:

1. Start by determining what concentration (molarity) and volume of Tris buffer you want to make. For example, Tris buffer solution used for saline varies from 10 to 100 mM. Once you have decided what you are making, calculate the number of moles of Tris that are required by multiplying the molar concentration of buffer by the volume of the buffer that is being made. (moles of Tris = mol/L x L)

2. Next, determine how many grams of Tris this is by multiplying the number of moles by the molecular weight of Tris (121.14 g/mol). grams of Tris = (moles) x (121.14 g/mol)

3. Dissolve the Tris into the distilled deionized water, 1/3 to 1/2 of your desired final volume.

4. Mix in HCl (e.g., 1M HCl) until the pH meter gives you the desired pH for your Tris buffer solution.

5. Dilute the buffer with water to reach the desired final volume of solution.

Once the solution has been prepared, it can be stored for months in a sterile location at room temperature. Tris buffer solution's long shelf life is possible because the solution does not contain any proteins.

Ref: https://www.thoughtco.com/how-to-make-tris-buffer-solution-603668