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application information | ||
recommended dilution | 1 : 1000 with standard ECL (WB) | |
expected | apparent MW | 115.7 kDa | |
confirmed reactivity | Arabidopsis thaliana | |
predicted reactivity | Brassica napus, Eucalyptus grandis, Populus sp. | |
not reactive in | no confirmed exceptions from predicted reactivity are currently known | |
additional information | to be added when available | |
selected references | to be added when available, antibody released in September 2013. |
500 mg of Col-0 WT Arabidopsis thaliana stem powder extracted by boiling in 2 mL of 3% SDS loading buffer + 100 mM DTT at 95C for 10 min. Extract was spun at max speed to remove debris and supernatant was taken as crude extract. 25 µL of this was loaded on a 4-15% gel run for 50 min, 150v. . Blots were blocked with 5 % milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500 over night at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602 from Agrisera) diluted to 1:5000 in for 2h at RT with agitation. The blot was washed as above and developed for 5 min with Pierce SuperSignal West Pico kit according to the manufacturer's instructions. Exposure time was 10 seconds. Courtesy of Dr. Manoj Kumar, University of Manchester, UK |