MIP1 | Aquaporin, glycerol transport activity
AS15 2826 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
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31.5 | 43 kDa
10 µg of total protein from 15ml cell suspension of Chlamydomonas reinhardtii CC3395, with a cell density of (~ 10 times 8) extracted with SDS buffer supplememnted with Protease inhibitor (cOmplete Ultra tablets, EDTA free, Roche, Mannheim), after removal of the cell debris via centrifugation, DTT was added (100mM final concentration) and were separated on 12 % SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with RotiBlock-Solution (Roth, Darmstadt) for 1h at room temperature (RT) or at 4°C ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10,000 in TBS with 2% milk powder for 1h at RT with agitation. The antibody solution was decanted and the blot was washed 3 times for 10 minutes each with TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Sigma, St.Luis, MO) diluted to 1:80 000 in TBS tith 5% milk powder for 1h at RT with agitation. The blot was washed as above and developed for 1-2 min with ECL according to the manufacturer's instructions. Exposure time was 40 seconds.
Courtesy of Dr. Karin Komsic-Buchmann and Prof. Dr. Burkhard Becker, University of Cologne, Germany
As MIP1 is a membrane protein please use a high redox potential in your lysis buffer.
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