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PHEM Buffer recipe

PHEM buffer has been used primarily for tissues and cell cultures being processed for immunocytochemical studies.  Most antigens, especially intra cellular ones, stain better using PHEM than those processed using PBS, it also has a more limited effect on biochemical reactions and enzymes. Very good preservation of cellular ultrastructure, notably microtubules, is also an advantage of PHEM use.


  1. Schliwa, Manfred, and Jonathan Van Blerkom. “Structural interaction of cytoskeletal components.” The Journal of Cell Biology 90.1 (1981): 222-235.
  2. Jacqueline Montanaro: Improved ultrastructure of marine invertebrates using non-toxic buffers

Preparation recipe for 500ml of Buffer PHEM at 0.2M

Dissolve in 400 ml distilled  water this reagents powder in following order:

Reageant powder Number C.A.S weight gram/ Molarity Ref. em-grade
HEPES 7365-45-9 0.598 gr.  /  5mM. GP19
EGTA 67-42-5 0.380 gr. /  2mM GP27
MgSO4 , 7H2O 10034-99-8 0.098 gr /  0,8mM GP28
P.I.P.E.S. 5625-37-6 1.814 gr /  12mM GP21
WATER  D. Make up to 500ml final volume and adjust to the desired pH with NaOH

Filter the solution to 0.22µ.

We also offer our ready-to-use PHEM buffer available on our shop: PHEM Buffer

Ref. : GT140000