Cyt c | Cytochrome c

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana cytochrome c protein sequence, UniProt:D7KMK0 (C-1) D7LY03 (C-2), TAIR: At1g22840 (Cytc1) and At4g10040 (Cytc2)

12.5 | 14 kDa (for Arabidopsis thaliana)

cytc1 and cytc2 from following species: A. theoprasi, Brassica napus, Brassica oleracea, Cannabis sativa, 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.

Arabidopsis thaliana CytC6

Application example

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 chemiluminescent detection reagent. Chemiluminescence was detected by BioSpectrum® Imaging System (UVP). Exposure time was 5 seconds.

Courtesy Dr. Janusz Piechota, Wrocław University, Poland

The presence of cytochrome c in the cysotol is a marker of PCD (programmed cell death).

Cytochrome c is located in inner mitochondrial membrane. It is a small heme protein which, unlike other cytochromes, is highly soluble. This protein is an essential component of the electron transport chain, where it undergoes oxidation and reduction without binding oxygen.