Recipes of Commonly Used Buffer Containing Guanidine Hydrochloride

As we all know, guanidine hydrochloride is often used as an intermediate in medicines, pesticides, dyes and other organic compounds, it is also an important raw material for sulfonamides and folic acid; it can be used as a strong denaturants in the extraction of total RNA from cells. At the same time, guanidine hydrochloride is often used in various buffers for biological experiments. Let us take a look at the following recipes.


1. Guanidine Hydrochloride (GuHCl) Buffer

• 6 M guanidine hydrochloride
• 8 mM imidazole
• 50 mM NaH2PO4
• 50 mM Tris

Adjust pH to 8.0. Prepare within a few days of the experiment and store at room temperature (20~25°C).


2. Denaturing buffer containing guanidine hydrochloride

• 6 M Guanidine hydrochloride
• 5 mM DTT
• 50 mM Tris-Cl (pH 7.5)

Warm the solution to 40ºC to dissolve the guanidine HCl. Adjust the pH to 7.5 if necessary.


3. Hydroxylamine cleavage buffer
Final concentrations: 2M hydroxylamine hydrochloride, 4.3M guanidine hydrochloride, 0.2 M potassium carbonate (pH 9.0)

• Hydroxylamine hydrochloride (Analar grade)
• Guanidine hydrochloride (6 M aqueous), prechilled
• Potassium carbonate (1 M)
• NaOH (50% w/v)

Dissolve 3.5g of hydroxylamine hydrochloride in 12mL of prechilled 6M aqueous guanidine hydrochloride (use an ice bath). Add 2mL of 50% (w/v) NaOH slowly with vigorous stirring (magnetic stir bar) followed by 5mL of 1M potassium carbonate. Adjust the pH to 9.0 with 50% NaOH, and then adjust the volume to 25 ml with 6 M guanidine hydrochloride.

The inclusion of guanidine hydrochloride is reported to improve the efficiency of Asn-Gly cleavage.


4. Solubilization buffer

• 8 M guanidine hydrochloride
• 50 mM Tris-Cl (pH 7.5)

or

• 10 mM DTT (dithiothreitol)
• 8 M guanidine hydrochloride
• 50 mM Tris-Cl (pH 8.5)

5. Denaturing buffer for Ellman’s test

• Guanidine hydrochloride (6 M)
• Na2HPO4 (0.1 M, pH 8.0)

6. SEC column buffers

• 50 mM Sodium phosphate, containing 150 mM NaCl (pH 7.0)
• Denaturing buffer containing urea
• Denaturing buffer containing guanidine hydrochloride
• Peptide elution buffers

Edited by Suzhou Yacoo Science Co., Ltd.