IGEM:Harvard/2006/Xuetal

From Xu et al 2004, they have anti-KaiC polyclonal antibodies which they used to do a phosphorylated KaiC assay; we can use this method to analyse KaiC fluctuation.

From the paper: KaiC Phosphorylation Assay. Cyanobacterial cultures were grown to an OD750 of 0.2. After a 12-h dark pulse, the cultures were treated with 10�MIPTGfor 3 h at 30°C in light with air bubbling, and�40 ml of the cultures were collected for preparation of total extracts. Immunoblot analysis for KaiC was performed on 5 �g of total proteins per lane according to the previous description (20), except that we used a highly specific mouse polyclonal antibody to KaiC and a SuperSignalWest Pico Chemiluminescent Substrate (Pierce). The phosphorylated KaiC signals were quantified as before (16) and analyzed by Student’s t test.