Quantitative rapid separation of histones on cellogel strips using dansyl chloride
✍ Scribed by M.O. Creighton; J.R. Trevithick
- Book ID
- 102983127
- Publisher
- Elsevier Science
- Year
- 1972
- Tongue
- English
- Weight
- 963 KB
- Volume
- 50
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Quantitative analysis of small quantities of proteins has often been impossible because of the lack of sensitivity or unsuitability of current methods for estimating proteins. Improvements in technique which permit the analysis of very small amounts of protein are thus very useful.
Since histones contain scant amounts of aromatic residues, their analysis presents a particular challenge. Wray and Stubblefield (1) recently reported a highly sensitive detection method for histones in acrylamide gels using a modification of the method of Sung and Smithies (2), which detected submicrogram amounts of histones in polyacrylamide gels; however, during the process of destaining the histone bands were distorted, making it difficult to estimate them accurately. Another disadvantage of the polyacrylamide electrophoresis of histones in general is that, although very good separations are possible, the time required for separation by the high-resolution method of Panyim and Chalkley (3) and the subsequent staining of the gel might be more than 24 hr altogether.
For these reasons we investigated both a more rapid method for separation of the bands of histones using electrophoresis on cellogel strips and an improved method of detection making use of fluorescent reagents. While this work was in progress, a report appeared of the separation of histones by cellogel electrophoresis by Machicao and Sonnenblicher (4) using conditions which differed substantially from ours.