Cyt c | Cytochrome c
AS08 343A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Lupinus luteus | Marker of PCD
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1: 5000 (WB)
|Expected | apparent MW||
12.5 | 14 kDa for A. thaliana
|Confirmed reactivity||Arabidopsis 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, Nicotiana tabacum, Oryza 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||Schimmeyer et al. (2016). L-Galactono-1,4-lactone dehydrogenase is an assembly factor of the membrane arm of mitochondrial complex I in Arabidopsis. Plant Mol Biol. 2016 Jan;90(1-2):117-26. doi: 10.1007/s11103-015-0400-4. Epub 2015 Oct 31.
Li et al. (2016). Characterization of a novel β-barrel protein (AtOM47) from the mitochondrial outer membrane of Arabidopsis thaliana. J Exp Bot. 2016 Nov;67(21):6061-6075. Epub 2016 Oct 6.
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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