PBST Buffer (PBS with Tween 20)
PBST or PBS-T (phosphate-buffered saline with Tween 20) is PBS buffer with a detergent such as Tween 20 or Triton X-100 (CSH Protocols recommends Tween 20). Detergent concentration can vary depends on requirement but generally vary from 0.05 – 2% (CSHL Protocols recommends 0.05). We are giving composition based on CSHL protocols in the tabular form.
Used as a washing solution for:
- Western blot membranes.
- PFA (Paraformaldehyde) fixed samples.
- Microtiter plate wells in ELISA assays.
Keep pH at 7.4 with HCl and NaOH.
To prepare 0.05% (V/V) PBS-T buffer
|1X PBS 500 mL||1X PBS 1L||10X PBS 500 mL||10X PBS 1L|
|Tween 20 to be added||25 µl||50 µl||250 µl||500 µl|
- 1X PBS (Click here for composition).
- Tween 20.
Material and instruments required
- Pipettes and tips.
- Glassware for mixing
- Measuring cylinder
- Make 1X PBS buffer
- Add Tween-20 as desired concentration
- Swirl slowly until dissolved
Store at room temperature. Keeping in lower temperature might lead to crystallization.
Tween 20 and Triton X-100 are viscous liquids. You will have to cut the narrow end of the pipette tip for easy aspiration.
If you find pipetting still difficult, weigh the detergent (consider density for determining volume) and add PBS to it.
Detergents make froth upon agitation. Swirl gently to avoid froth.