Immunofluorescent analysis of Phalloidin (orange) and Cytochrome c (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Cytochrome c monoclonal antibody (Product # MA5-11823) at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 550 Phalloidin (Product # 21835) at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst (Product # 62249) at a concentration of 1ug/ml for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.
|Tested species reactivity||Bovine, Dog, Horse, Human, Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG2a, kappa|
|Immunogen||Equine cytochrome c|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunofluorescence (IF)||1:100 - 1:500|
|Immunoprecipitation (IP)||2 µg/ml|
* 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.
|Immunocytochemistry (ICC)||See 1 publications below|
MA5-11823 targets Cytochrome c in ELISA, FACS, immunofluorescence and immunoprecipitation applications and shows reactivity with Bovine, Canine, Equine, Mouse and Human samples. MA5-11823 is not suitable for Western blotting applications.
This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Cytochrome c is an electron transporting protein that resides within the inter-membrane space of the mitochondria, where it plays a critical role in the process of oxidative phosphorylation and production of cellular ATP. An increasing amount of interest has been directed toward the role which cytocrome c has been demonstrated to play in apoptotic processes. Following exposure to apoptotic stimuli, cytochrome c is rapidly released from the mitochondria into the cytosol, an event which may be required for the completion of apoptosis in some systems. Cytosolic cytochrome c functions in the activation of caspase-3, and ICE family molecule that is a key effector of apoptosis.
IP-MS enrichment of CYCS (LFQ intensity): CYCS was enriched 1894-fold from MCF7 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and CYCS antibody (Part No. MA5-11823). See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC).
MA5-11823 was used in immunocytochemistry to evaluate the toxicological effects of harmful and potentially harmful constituents (HPHC) by utilizing a high content screening analysis
|Marescotti D,Gonzalez Suarez I,Acali S,Johne S,Laurent A,Frentzel S,Hoeng J,Peitsch MC||Journal of visualized experiments : JoVE (null:null)||2016|