AS12 1842 | clonality: polyclonal | host: rabbit | reactivity: Solanum tuberosum
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|Recommended dilution||1 : 100 (IL), 1 : 2000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Solanum tuberosum
|Predicted reactivity||Solanum tuberosum|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Additional information||Imunolocalization analyses were performed on 10-mm-thick bud meristem sections cut by microtome following the protocol described by Paciorek et al. (2006).
The following antibodies and dilutions were used: anti- Patatin (1:100) (Agrisera, Sweden) and Alexa Fluor® 488 Goat Anti-Rabbit IgG (H+L) secondary antibody (1:100) (Molecular probes).
Immunofluorescence was observed with an IX81/FV500 confocal laser-scanning microscope (Olympus) equipped with a 488-nm argon ion laser and a 405-nm diode laser.
to be added when available, antibody released in October 2012.
75 µg of total protein from Solanum tuberosum v. Nicola extracted with 20 mM TRIS PH-8.5, 10 mM thiourea,10mM CaCl2, 5mM DTT, 1 mM PMSF, 1%PVPP were separated on 4-20 % SDS-PAGE and blotted 1.5h to PVDF. Blots were blocked with 5% milk powder in T-TBS (2.5gr of powder in 50 ml T-TBS) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 for overnight at 4°C with agitation. The antibody solution was decanted and the blot was washed 6 times for 5 min. 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 1:10 000 in blocking solution for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 30 seconds.
Courtesy of Dr. Paula Teper-Bamnolker and Dr. Dani Eshel, The Volcani Center, Israel
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