Fig. 4. Immunoprecipitation of CKI1-GFP in the lysate from an Arabidopsis suspension cell culture. (A) 10 mg of total solubilized proteins (input) were purified using GFP-Trap (IP) from the Pro35S:CKI1-GFP suspension cell culture and after separation on a 7% SDS PAGE gel, electro-blotted onto a PVDF membrane and detected using oiCKI1RD PAbs (1:3,000; left) or oiGFP antibodies (1:1,000; right). (B) 5.5 mg of total solubilized proteins (input) from the Pro35S:CKI1-GFP suspension cell culture were precipitated with oiCKI1RD PAbs bound to A/G agarose beads (IP). The eluted proteins were separated on a 7% SDS–PAGE gel, electro-blotted onto a PVDF membrane and probed with oiCKI1RD PAbs (1:3,000; left) or oiGFP MAb (1:1,000; right). Wt, wild-type Landsberg erecta; M, molecular weight marker; IP, immunoprecipitation. Immunoprecipitations are not quantitative.

Published as part of Borkovcová, Petra, Pekárová, Blanka, Válková, Martina, Dopitová, Radka, Brzobohatý, Břetislav, Janda, Lubomír & Hejátko, Jan, 2014, Antibodies against CKI1 , a receiver domain of the sensor histidine kinase in Arabidopsis thaliana: From antigen preparation to in planta immunolocalization, pp. 6-15 in Phytochemistry 100 on page 10, DOI: 10.1016/j.phytochem.2014.01.007, http://zenodo.org/record/10480315