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Strains with or without Myc-tagged orf19.3456 and HA-tagged orf19.193 were grown to log phase in YPD at 30°C. Protein extracts were prepared as described in materials and methods and immunoprecipitated with an anti-Myc antibody. The immunoprecipitated orf19.3456 -encoded kinase was detected by Western blotting with an anti-Myc antibody, and the co-immunoprecipitated orf19.193 -encoded protein with an anti-HA antibody (top panels, IP). Tagged proteins in the input samples were detected with anti-HA and anti-Myc antibodies (bottom panels, input). Detection of tubulin with <t>an</t> <t>anti-tubulin</t> antibody served as loading control for the input samples. Both independently generated series of strains were used for the experiment.
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Strains with or without Myc-tagged orf19.3456 and HA-tagged orf19.193 were grown to log phase in YPD at 30°C. Protein extracts were prepared as described in materials and methods and immunoprecipitated with an anti-Myc antibody. The immunoprecipitated orf19.3456 -encoded kinase was detected by Western blotting with an anti-Myc antibody, and the co-immunoprecipitated orf19.193 -encoded protein with an anti-HA antibody (top panels, IP). Tagged proteins in the input samples were detected with anti-HA and anti-Myc antibodies (bottom panels, input). Detection of tubulin with an anti-tubulin antibody served as loading control for the input samples. Both independently generated series of strains were used for the experiment.

Journal: PLOS Genetics

Article Title: Inducible gene deletion reveals essentiality of protein kinases and a septation initiation network in Candida albicans

doi: 10.1371/journal.pgen.1012118

Figure Lengend Snippet: Strains with or without Myc-tagged orf19.3456 and HA-tagged orf19.193 were grown to log phase in YPD at 30°C. Protein extracts were prepared as described in materials and methods and immunoprecipitated with an anti-Myc antibody. The immunoprecipitated orf19.3456 -encoded kinase was detected by Western blotting with an anti-Myc antibody, and the co-immunoprecipitated orf19.193 -encoded protein with an anti-HA antibody (top panels, IP). Tagged proteins in the input samples were detected with anti-HA and anti-Myc antibodies (bottom panels, input). Detection of tubulin with an anti-tubulin antibody served as loading control for the input samples. Both independently generated series of strains were used for the experiment.

Article Snippet: For the detection of tubulin, membranes were blocked with 5% milk in TBST and incubated overnight at 4°C with rat anti-tubulin alpha antibody MCA 78G (Bio-Rad), washed with TBST, and then incubated with rabbit anti-rat HRP-conjugated antibody STAR21B (Bio-Rad).

Techniques: Immunoprecipitation, Western Blot, Control, Generated