PBST Buffer (PBS with Tween 20)

PBST or PBS-T (phosphate buffered saline with Tween 20) is PBS buffer with 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% (CSHP recommends 0.05). We are giving composition based on CSHL protocols in the tabular form.

Applications

Used as washing solution for:

  1. Western blot membranes.
  2. PFA (Paraformaldehyde) fixed samples.
  3. Microtiter plate wells in ELISA assays.
  4.  Immunohistology.

pH

Keep pH at 7.4 with HCl and NaOH.

Composition

PBS-T composition
0.05% (V/V) Tween 20. pH 7.4.
PBS-T composition
0.05% (V/V) Tween 20. pH 7.4.
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

Reagents required

  1. 1X PBS (Click here for composition).
  2. Tween 20.

Material and instruments required

  1. Pipettes and tips.
  2. Glassware for mixing
  3. Measuring cylinder

Procedure

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

Storage

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

Caution

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.

References

  1. Cold Spring Harbor Protocols
  2. Openwetware
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