Cyt c | cytochrome c
AS08 343A | clonality:polyclonal | host: rabbit | reactivity: Arabidopsis thaliana, Lupinus luteus| marker of PCD
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1: 5000 with standard ECL (WB)
|Expected | apparent MW||
12,5 | 14 kDa for A. thaliana
cytc1 and cytc2 from following species: A. theoprasi, B. napus, B. oleracea, C. maxima, Ch. reinhardtii (peptide target partially conserved), L. esculentum, Lupinus luteus, M. truncatula, O. sativa, Ostreococcus (peptide target partially conserved), P. aurea, Physcomitrella patens, R. comunis, S. nigra, V. vinifera
|Not reactive in||
Arabidopsis thaliana CytC6
The presence of cytochrome c in the cysotol is a marker of PCD (programmed cell death).
|Selected references||to be added when available, antibody release in November 2014|
Mitochondrial proteins (15 ug) from Arabidopsis thaliana mitochondria was separated on 16% acrilamide gel and electrophoresis prepared according to Schägger and von Jagov (Anl. Biochem., 1987, 166:368-379). After running the gel, proteins were transferred to PVDF membrane using wet transfer (Roti®-Blot 2, Roth). Transfer was checked by Ponceau S staining. Blot was destained by several quick washings in distilled water and 1 washing in 1X TBS (10 mM T pH 7.5, 150 mM NaCl) (10-15 min.).Blot was blocked by 1.5 hour in 5% milk in TBST (1X TBS, 0,1% Tween 20) After blocking blot was washed quickly twice in TBST and incubated 2 hours with primary antibody (dilution 1: 1000) in TBST. Washing: two quick washings in TBST and 3 x 10 min. washings in TBST. Then blot was incubated 45-60 min. with a secondary anti-rabbit antibodies conjugated to peroxidase (Agrisera AB, dilution 1:10 000, AS09 602) in TBST. Washing: as above. After washing blot was incubated 1-2 min. in ECL solution (NOWA Kit, Thermo Scientific). Chemiluminescence was detected by BioSpectrum® Imaging System (UVP). Exposure time was 5 seconds.
Courtesy Dr. Janusz Piechota, Wrocław University, Poland
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