STN7 | Serine/threonine-protein kinase STN7 (chloroplastic)
AS16 4098 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Nicotiana tabacum
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Lyophilized antibody can be stored at -20°C for up to 3 years. Re-constituted antibody can be stored at 4°C for several days to weeks.
Once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
63.2 | 44 kDa (on urea gel), 55 kDa (no urea)
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Isolated thylakoids from Arabidopsis thaliana leaves were extracted with grinding buf fer containing 300mM Sucrose, 50 mM Hepes-NaOH pH 7,4, 5 mM MgCl , 1 mM Na-EDT A containing freshly made 10 mM NaF. Then the 2 chloroplast pellet was dissolved with shock buf fer containing, 5 mM Sucrose, 10 mM Hepes-NaOH pH 7,4, 5 mM MgCl containing freshly made 2 NaF (10 mM), whereafter pellet containing thylakoid fraction was carefully again dissolved in to the storage buf fer containing 100mM sucrose, 10 mM Hepes-NaOH pH 7,4, 10 mM MgCl in the presence of freshly made 10 mM 2 NaF . Protein amounts loaded according to PORRA corresponding to 1, 2, or 4μg of Chlorophyll, and denatured with Laemmli buf fer at C or 65C for 5 min. Proteins were then further separated on 15 % Acryl Amide containing 6M Urea with SDS-P AGE, blotted 1h to PVDF membrane using semi-dry transfer (Hoefer TE77X) followed by rinsing in 1xTBS for 2 min. Membranes were then blocked with 4% Milk in 1XTTBS for 2 hrs at room temperature (R T) with agitation. Blot was then incubated overnight in af finity purified Stn7 primary antibody (Agrisera) at a dilution of 1:1500 for 18hrs at +4C with slow agitation in 1% Milk, 1XTTBS. The antibody solution was collected, and the blot was rinsed 5 minutes 3 times each with 1x T-TBS at RT with agitation. Blot was incubated with HRP conjugated conjugated goat anti-rabbit IgG secondary antibody (Agrisera, AS09 602) diluted to 1:25 000 in 1% Milk, 1XTTBS for 2hrs at R T with agitation. The blot was washed 10 min in 1X TTBS, with additional washes of 4X4 min in 1XTBS. Blot was then incubated in ECL solution for 5 min. Film was exposured for 2minutes.
Courtesy of Dr. Jouni Tivola, University of Turku, Finland
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