Anti-MKK1 | Mitogen-activated protein kinase kinase 1

Hordeum vulgare, Musa sp., Oryza sativa, Phaseolus vulgaris, Physcomitrium patens, Solanum tuberosum, Ricinus communis,Triticum aestivum, Zea mays 

Ostreococcus tauri

application example

10 µg of total protein from Arabidopsis thaliala whole seedlings, various cellular fractions, extracted with (see below*) were separated on 15 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with 5% Non fat dairy milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 3000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was washed four times for 5 min inPBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:15 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 180 seconds.

*Following is the protein extraction buffer composition that was used to extract the total protein from the whole seedlings:50mM Tris, 4% SDS, 1% PVPP , 5% Glycerol, 1mM PMSF, 2mM Pefabloc.

Courtesy of Anshika Jain and Erin Connolly, University of South Carolina, USA

to be added when available

antibody is recognizing a chloroplastic form of plant ferritin