PBST Buffer (PBS with Tween 20)

Table Of Contents


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.
  • Immunohistology.


1X PBS 500 mL1X PBS 1L10X PBS 500 mL10X PBS 1L
Tween 20 to be added25 µl50 µl250 µl500 µl

Table 1. Preparation of 0.05% (V/V) PBS-T buffer.


Keep pH at 7.4 with HCl and NaOH.


  1. Make 1X PBS buffer.
  2. Add Tween-20 as desired concentration.
  3. Swirl slowly until dissolved.


Store at room temperature. Keeping in lower temperature might lead to crystallization.

Points to be noted

  • 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.
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