A rapid paper chromatographic assay for protein kinase

✍ Scribed by Heng-Chun Li; David A. Felmly

• Publisher: Elsevier Science
• Year: 1973
• Tongue: English
• Weight: 275 KB
• Volume: 52
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(73)90352-7

✦ Synopsis

SHORT COMMUNICATIONS

A Rapid

Paper Chrokatographic Assay for Protein Kinase

This communication describes a simple, rapid, and effective paper chromatographic method for separating the trichloroacetic acid (TCA) insoluble phosphorylated histone from its substrate Y-~~P-ATP in the protein kinase reaction. This method can be applied to the separation of TCA precipitable macromolecules (e.g., DNA, RNA, and protein) from their respective small molecule precursors (e.g., nucleotides and amino acids).

A sheet of 57.2 X 45.4-cm Whatman 31 ET chromatographic paper is cut into four sheets of 28.6 X 22.7 cm. Each sheet is scored off with pencil and, as shown in Fig. 1, fourteen 2 X 2-cm sample squares can be accommodated on a paper of this size. The paper is folded sharply along the line 4 cm from the edge (upper dotted line shown in Fig. 1). The sample squares are 1 cm from the fold. The part above the fold is immersed in the solvent. The lower dotted line, which is 10 cm below the lower bound-FIG. 1. Dimensions of the chromatography paper. The sample aliquots are applied in the center of each 2 X 2-cm square. The arrow indicates the direction of the solvent flow. 300