A Colorimetric Assay for Heme in Biological Samples Using 96-Well Plates

✍ Scribed by A.V. Pandey; S.K. Joshi; B.L. Tekwani; V.S. Chauhan

• Publisher: Elsevier Science
• Year: 1999
• Tongue: English
• Weight: 48 KB
• Volume: 268
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1998.2997

✦ Synopsis

ers such as piperazine diacrylamide in place of bisacrylamide. In general, if gel conditions other than those described in this paper are to be employed, an experiment analogous to that shown in Fig. 2a should be carried out to determine the optimal PEG concentration for those specific conditions.

In summary, to prevent distortion of gel wells during preelectrophoresis of acid-urea or Triton-acid-urea slab gels, proceed as follows: Before preelectrophoresis, fill the gel wells with a solution containing the appropriate concentration of PEG 8000 (10% (w/v) for gels containing 15% acrylamide and 0.1% bisacrylamide) and the same concentrations of urea, acetic acid, and Triton as are present in the gel. After preelectrophoresis, rinse out the wells with water before filling them with electrode buffer and loading samples. This procedure greatly improves the quality, reproducibility, and convenience of protein separations in acid-urea slab gels.

Acknowledgments. We are grateful to J. A. Brown and L. C. Lutter for discussion and to the Faculty Development Board of the University of Wisconsin Oshkosh for support.