Use of phospho-cellulose paper disks for the assay of histone acetyltransferase

✍ Scribed by Kentaro Horiuchi; Daisaburo Fujimoto

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 320 KB
• Volume: 69
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90151-7

✦ Synopsis

A modified method for the assay of histone acetyltransferase is presented. Previously reported methods depended upon the determination of the incorporation of radioactivity from [Wlacetyl coenzyme A into trichloroacetic acid (TCA)-precipitable material. However, as shown in this paper, [14C]acetylated histone cannot be precipitated quantitatively by TCA. In the method described in this paper, phospho-cellulose (P-cellulose) paper disks are used as an adsorbent for ['%]acetylated histone and 0.05 M carbonate buffer, pH 9.2, is used as a washing medium. This P-cellulose disk method allows more quantitative determination of [Wlacetylated histone than the TCA-precipitation methods.

METHODS

Preparation of Histone

Whole histone was prepared from calf thymus by extracting with 0.25 N H,SO, as described by Murray (8). f2a2 Histone was prepared 491