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Electron Microscopy Sciences

Technical Data Sheets

PHEM Buffer

EMS #11162, 11163

Introduction

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.

Desired Fixative

200 ml of 2.5% Glutaraldehyde, 1.5% Paraformaldehyde in 0.1M PHEM buffer.

Glutaraldehyde (Cat#1 6320) 50% x V1 = 2.5% x 200 ml V1 = 10 ml of 50% EM Grade Glutaraldehyde
Paraformaldehyde (Cat# 15710-S) 16% x V1 = 1.5% x 200ml V1 = 18.75 ml of 16% Paraformaldehyde
0.2 M PHEM (Cat# 11162) 0.2 M x V1 = 0.1M x 200 ml

V1= 100ml of 0.02M PHEM Buffer

Adjust to desired pH. QS with distilled water to 200 ml.

References

  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
    Academic Editor: James Reimer, published online March 31, 2016

Product Information

PHEM Buffer