How do you make sodium deoxycholate solution?

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). Store the buffer at 4°C.

How do you make a 10 sodium deoxycholate solution?

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.

How do you make a 1X TBS buffer?

To prepare, dissolve 6.05 g Tris and 8.76 g NaCl in 800 mL of H2O. Adjust pH to 7.5 with 1 M HCl and make volume up to 1 L with H2O. TBS is stable at 4°C for 3 mo.

What is sodium deoxycholate for?

Sodium deoxycholate (deoxycholic acid) is a water-soluble, bile-acid, ionic detergent commonly used in protein methods. Sodium deoxycholate is the detergent recommended for stripping endotoxin (Lipopolysaccharide or LPS) from immobilized Polymyxin B columns.

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 the pH of Tris?

Making a Tris Buffer Tris buffer is a good choice for most biological systems because it has a pKa of approximately 8.1 at 25°C, making it an effective buffer in the range of pH 7–9.

What is TE buffer?

TE Buffer, 1X, Molecular Grade (pH 8.0), is a buffer composed of 10mM Tris-HCl containing 1mM EDTA•Na2. Properties: pH at 25°C: 7.9–8.1.

How do I convert 1X TBS to 20X?

To make a 1X PBS solution dilute concentrate 20X with distilled water. Measure and pour appropriate volume of 20X PBS concentrate into a mixing flask and add DI water to final volume. Stir briefly. The 1X solution should be pH 7.6 ± 0.2.

How do I convert 1X TBS to 10X?

TBS 10x (concentrated Tris-buffered saline) For a 1x solution, mix 1 part of the 10x solution with 9 parts distilled water and adjust pH to 7.6 again. The final molar concentrations of the 1x solution are 20 mM Tris and 150 mM NaCl.

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.

What does sodium deoxycholate do in RIPA buffer?

The following RIPA Lysis Buffer components have the following effects: Tris-HCl is a buffering agent prevents protein denaturation, NaCl is a salt that prevents non-specific protein aggregation, NP-40 is a non-ionic detergent to extract proteins; Na-deoxycholate and SDS are ionic detergents to extract proteins; and …

Is Tris a base or acid?

What is Tris HCl. Tris HCL is a buffering agent (acidic buffer) commonly used by molecular biologists to adjust the pH of a solution or stabilize the pH. Commercially available Tris HCl is Tris with HCl added.

Is there a 10% sodium deoxycholate stock solution?

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 H 2O and then add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the total volume to 50 ml). Store the buffer at 4°C. Nonidet-P40 (NP-40) buffer

How to make sodium deoxycholate with ultrapure water?

To each tube, add 900 μL of ultrapure water, then 200 μL of sodium deoxycholate reagent, mix, and let stand at room temperature for 10 min. 4. Add 200 μL of the TCA reagent, thoroughly mix, and then centrifuge for 15 min at 1000–3000 × g. 5. After centrifugation, gently remove supernatant.

What is the molecular weight of sodium deoxycholate?

Thermo Scientific Sodium Deoxycholate is an ionic detergent that is especially useful for disrupting and dissociating protein interactions. • Molecular Weight: 414.6g • Molecular Weight: 414.6g • Absorbance (1% Detergent Solution): 340nm <0.02; 280nm <0.04; 260nm <0.06 Sodium deoxycholate (deoxycholic acid)…

How to make sodium deoxycholate in a centrifuge?

Pipette 100 μL of chitosan solution into centrifuge or microcentrifuge (at least 2 mL vol) tubes in triplicate. 3. To each tube, add 900 μL of ultrapure water, then 200 μL of sodium deoxycholate reagent, mix, and let stand at room temperature for 10 min. 4.