how to make 100ml of TBST buffer from 1X TBS (tris buffered saline solution) and 0.1% tween-20

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how to make 100ml of TBST buffer from 1X TBS (tris buffered saline solution) and 0.1% tween-20. I am just confused how to do the calculations when it is 1X TBS and not 10X TBS. Please show calculations

The correct answer and explanation is :

To make 100 mL of TBST buffer from 1X TBS and 0.1% Tween-20, follow these calculations:

Step 1: Understanding the components

  • 1X TBS is already at the correct concentration, so you do not need to dilute it further.
  • Tween-20 needs to be added to a final concentration of 0.1% (v/v).

Step 2: Calculate Tween-20 volume

  • 0.1% Tween-20 means 0.1 mL of Tween-20 per 100 mL of solution.
  • Convert to microliters:
    [
    0.1 \text{ mL} = 100 \text{ µL}
    ]

Step 3: Prepare the solution

  1. Measure 100 mL of 1X TBS.
  2. Add 100 µL of Tween-20.
  3. Mix well by gentle stirring or inversion.

Explanation (300 words)

TBST (Tris-Buffered Saline with Tween-20) is a commonly used buffer in biochemical applications, particularly in Western blotting and ELISA to wash membranes and prevent non-specific binding. The key components of TBST are TBS (Tris-buffered saline), which maintains pH stability, and Tween-20, a detergent that helps reduce background noise by preventing non-specific protein binding.

In this case, we are starting with 1X TBS, which means it is already at its working concentration. Unlike working with 10X TBS, where dilution calculations are required, 1X TBS does not require further dilution. Our only step is to add Tween-20.

Since Tween-20 is typically used at a 0.1% (v/v) concentration, we calculate its volume using the formula:
[
\text{Volume of Tween-20} = \text{Final Volume} \times \text{Desired Percentage}
]
[
= 100 \text{ mL} \times 0.1\% = 0.1 \text{ mL} = 100 \text{ µL}
]

Once the Tween-20 is added, the solution should be mixed gently to ensure even distribution. It is best to store TBST at 4°C if not used immediately.

This simple preparation ensures consistency and reproducibility in experiments where TBST is required as a washing buffer, improving the reliability of Western blot and ELISA results.

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