AHK2 | Histidine kinase 2
AS12 2113 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
|Recommended dilution||1 : 1000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Vitis vinifera|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
to be added when available, antibody released in July 2013.
The Arabidopsis thaliana wild-type (WS) and ahk2-2 T-DNA insertion mutant plants were grown on MS/2 media for six days. Tissue was collected, frozen in liquid nitrogen, ground in 3 volumes of 2x SDS-PAGE loading buffer and heated at 95 C for 5 min. After 10 min centrifugation at RT, 20 µl of the supernatant was loaded per lane on a 7.5% TGX gel (BioRad). After separation, proteins were blotted 1 hr onto a supported nitrocellulose membrane. Blots were blocked with 10% milk in PBS-T for 15 min at room temperature (RT) with agitation. Blot was incubated in the primary antibody diluted 1: 10, 000 in 1% milk in PBS-T for 16h at 4C with agitation. The antibody solution was decanted and the blot was washed 3 times for1 5 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 2000 in PBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5 min with SuperSignal West Femto Substrate according to the manufacturers instructions. Chemiluminescent signal was captured using ChemiDoc equipped with a CCD camera.
Courtesy of Dr. Jasmina Kurepa, University of Kentucky College of Agriculture, Food and Environment, USA
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