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Western blot analysis of cytochrome C was performed by loading 20 µg of MCF-7 (lane 1) and MDA-MB-231 (lane 2) cell lysates in RIPA buffer (Product # 89901) and Page Ruler Plus Protein Ladder (Product # 26620) onto a 4-8% sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE). Proteins were transferred to nitrocellulose membrane blocked in 5% milk /PBST for one hour at room temperature. Cytochrome C was detected using a cytochrome C monoclonal antibody (Product # 45-6100) at a dilution of 1:500 5% milk /PBST overnight at at 4°C on a rocking platform, followed by a Goat anti-Mouse IgG (H + L) -HRP secondary antibody at a dilution of 1:5,000 for one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34078) and Kodak M35 X-OMAT Automatic Processor. Data courtesy of Dr. Wei Xu at the University of Wisconsin, Madison, WI.
|Tested species reactivity||Human|
|Published species reactivity||Mouse|
|Host / Isotype||Mouse / IgG2a, kappa|
|Storage buffer||HEPES buffered saline|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||0.5-2 ug/ml|
|Immunofluorescence (IF)||Assay Dependent|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
45-6100 was used successfully in the western blot analysis of cytochrome C using human breast cancer cell line lysates.
This gene encodes cytochrome c, a component of the electron transport chain in mitochondria. The heme group of cytochrome c accepts electrons from the b-c1 complex and transfers electrons to the cytochrome oxidase complex. Cytochrome c is also involved in initiation of apoptosis. Upon release of cytochrome c to the cytoplasm, the protein binds apoptotic protease activating factor which activates the apoptotic initiator procaspase 9. Many cytochrome c pseudogenes exist, scattered throughout the human genome.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Specific acetylation of p53 by HDAC inhibition prevents DNA damage-induced apoptosis in neurons.
45-6100 was used in western blot to investigate the effect of p53 acetylation on DNA damage-induced neuronal apoptosis.
|Brochier C,Dennis G,Rivieccio MA,McLaughlin K,Coppola G,Ratan RR,Langley B||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:8621)||2013|
Mitochondrial DNA toxicity compromises mitochondrial dynamics and induces hippocampal antioxidant defenses.
45-6100 was used in immunohistochemistry - paraffin section to investigate the effects of mitochondrial DNA damage on hippocampal neurons.
|Lauritzen KH,Cheng C,Wiksen H,Bergersen LH,Klungland A||DNA repair (10:639)||2011|
CYC; cytochrome c; HCS; THC4
CYC; CYCS; HCS; THC4