Gelatin Zymography Reagent Recipes

Youssef Farhat, Written 6/1/13, Last updated 5/6/14

Download the PDF or Microsoft Word versions of these recipes

Introduction

Our Gelatin Zymography Protocol makes use of pre-made buffers and gels, but most of these can be home-made for a fraction of the cost.  Here are some recipes for analogous buffers that have been published in the literature [1], and the same article also describes how to make your own gelatin zymograms.

Disclaimer: These recipes are provided for the convenience of the reader and are not guaranteed to work.  I have not attempted to make these reagents on my own, so be sure to take any necessary safety precautions while preparing them and realize that they may not be compatible with or work as well as the commercial reagents used in our Gelatin Zymography Protocol.  Please let us know if you tried these recipes and tell us whether they worked for you!

Recipes

  • 4X Sample Buffer (Note: Our gelatin zymography protocol uses 2X sample buffer, so be sure to adjust accordingly and dilute samples 3:1 (vol:vol) with 4X sample buffer instead of 1:1 as should be done with 2X sample buffer)
    • 250 mM Tris-HCl, pH 6.8
    • 40% v/v glycerol
    • 8% w/v SDS
    • 0.01% w/v bromophenol blue
    • Aliquot and store at –20°C. Warm before use to dissolve SDS
  • 10X Running Buffer
    • 0.25 M Tris base (such as J.T. Baker, #X171)
    • 1.92 M glycine,
    • The pH should be 8.3 and does not require adjusting
  • 10X Renaturing Solution
    • 25% v/v Triton X-100 in deionized water
  • 10X Developing Buffer
    • 500 mM Tris-HCl, pH 7.8
    • 2 M NaCl
    • 50 mM CaCl2
    • 0.2% v/v Brij 35

References

[1]   Toth, M. and R. Fridman. Methods in molecular medicine, 2001. 57: p. 163-74.

Citing These Recipes

Farhat, Y. “Gelatin Zymography Reagent Recipes.” Protocol Place. Jun 2013. Accessed on mm/dd/yyyy. <http://protocol-place.com/>

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