TPL | Transcription factor TOPLESS (rabbit antibody)
AS16 3207 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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1 : 1000 (WB)
|Expected | apparent MW||124 kDa|
|Confirmed reactivity||Arabidopsis thaliana
|Predicted reactivity||Gossypium arboreum, Gossypium hirsutum, Noccaea caerulescens, Populus tomentosa, Theobroma cacao|
|Not reactive in||Zea mays|
To be added when available, antibody released in February 2018.
20 µg of total protein from Arabidopsis thaliana Col-0 (wt) (1), Zea mays leaf total protein extract (2), tpl/tpr1/tpr4 triple mutant in Col-0 background (3), Tpl mutant (N655127) (4), extracted with 1x LDS loading buffer to 50 mg tissue powder and proteins were denatured at 75°C for 10 min (1x LDS buffer: 10% glycerol, 250 mM Tris-HCl pH 8.5, 0.5 mM EDTA, 2% Lithium dodecyl sulfate, 0.005% Bromophenol blue) were separated on % SDS-PAGE and blotted 1h to nitrocellulose using Trans Blot Turbo system (BioRad). Blots were blocked with 5 % nonfat milk in TBS-T (100 mM Tris-HCl, 200 mM NaCl, 0.05% Tween 20) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 5 min 4 times in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed with SuperSignal West PICO Plus Chemiluminescent detection kit (Thermo Scientific). Exposure time was 12.6 seconds.
Peptide used to elicit TPL1 antibody is not conserved in TPL1 of Zea mays, which therefore serves as a negative control.
Courtesy Dr. Janos Bindics,Gregor Mendel Institute of Molecular Plant Biology, Vienna, Austria
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