Cyt c | Cytochrome c
AS08 343A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Lupinus luteus | Marker of PCD
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|Recommended dilution||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, Brassica napus, Brassica oleracea, C. maxima, Chlamydomonas reinhardtii (peptide target partially conserved), Lupinus luteus, Medicago truncatula, Nicotiana tabacum, Oryza sativa, Ostreococcus (peptide target partially conserved), P. aurea, Physcomitrella patens, Ricinus communis, S. nigra, Solanum lycopersivum, Vitis 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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