How do you make a 10% deoxycholate solution of sodium?

10% (w/v) sodium deoxycholate (Sigma, D6750-100G) Dissolve 5 g of sodium deoxycholate in 30 ml of H2O and add distilled H2O to 50 ml. Store at 4°C. 1 M sodium phosphate dibasic (Na2HPO4) Dissolve 7.098 g of Na2HPO4 in 30 ml of H2O and add distilled H2O to 50 ml. Store at room temperature.

What is Tris HCl used for?

Short for Tris (hydroxymethyl) aminomethane (THAM) hydrochloride, Tris HCl is an organic compound often used in buffer solutions such as TAE or TBE for electrophoresis gels. Tris is highly soluble in water and is useful in the pH range 7.0-9.0.

How would you prepare 10 mM Tris HCl pH 8?

To obtain a 10 mM Tris-HCl pH 7.4 solution, dilute 1 M Tris-HCl pH 7.4 1:100 with nuclease-free water. For example, add 1 mL of 1 M Tris-HCl pH 7.4 to 99 mL of nuclease-free water. Always add an acid to an aqueous solution; never add an aqueous solution to an acid.

How do you dilute sodium deoxycholate?

Add 5 mL of the sodium deoxycholate solution of 10% that is prepared separately in advance (the working concentration of sodium deoxycholate solution is 0.5%), and add 1 mL of NP-40. Mix them and add distilled water until the total volume is 100 mL….Preparation of Buffer for WB.

What is Tris buffer made of?

What Is Tris Buffer Solution? Tris is short for tris(hydroxymethyl) aminomethane, a chemical compound which is often used in saline because it is isotonic and non-toxic.

What is EDTA buffer?

EDTA (ethylenediaminetetraacetic acid) is a chelating agent that binds divalent metal ions such as calcium and magnesium. EDTA can be used to prevent degradation of DNA and RNA and to inactivate nucleases that require metal ions. EDTA can also be used to inactivate metal ion-requiring enzymes.

Why does SDS PAGE have two pH?

Gel Layers: It Takes Two The top (stacking) layer has a lower percentage of acrylamide and a lower pH (6.8) than the bottom (resolving) layer, which has more acrylamide and a higher pH (8.8). SDS PAGE is run in a discontinuous buffer system. The running buffer has different ions and a different pH than the gels.

What pH is Tris HCl?

Tris Buffer (Tris-HCl) pH 7.4, pH 8.0 and pH 8.3.

Does sodium deoxycholate denature protein?

Sodium deoxycholate and sodium cholate are bile salts detergents. They are both anionic detergents. Deoxycholate does denature proteins while cholate is a non-denaturing detergent.

How do you make Na deoxycholate?

The 10% sodium deoxycholate stock solution (5 g into 50 ml) must be protected from light. The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O and then add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the total volume to 50 ml).