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| application information | ||
| recommended dilution | 1 : 10 000 with standard ECL (WB) | |
| expected | apparent MW | 10.5 kDa | |
| confirmed reactivity | Arabidopsis thaliana, Chlamydomonas reinhardtii | |
| predicted reactivity | Glycine max, Solanum lycopersicum, Oryza sativa, Pinus sp., Ricinus communis, Micromonas sp., Ostreococcus lucimarinus, Volvox carteri | |
| not reactive in | no confirmed exceptions from predicted reactivity are currently known | |
| additional information | to be added when available | |
| selected references | Ramundo et al. (2013). Repression of Essential Chloroplast Genes Reveals New Signaling Pathways and Regulatory Feedback Loops in Chlamydomonas. The Plant Cell. | |
application example
application example
 | 10 µg of total protein from Chlamydomonas reinhardtii extracted with standard lysis buffer (Tris-HCl pH 6.8 50mM, 10mM EDTA, 2% SDS, Sigma protease inhibitors) were separated on 15% SDS-PAGE and blotted tonitrocellulose membrane using a wet transfer cell.Blot was blocked in TBS-T containing 5% non-fat dry milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 in TBS-T containing 5% non-fat dry milk for 1h at RT with agitation. The antibody solution was decanted and the blot was washed 3 times for 15 min in TBS-T containing 1% non-fat dry milk with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Promega) diluted to 1:0 000 in TBS-T containing 1% non-fat dry milk for 1h at RT with agitation. The blot was washed as above and developed for 1 min with ECL according to the manufacturers instructions. Exposure time was 30 seconds. |
