Immunoblot was carried out as described earlier (30 (link)). Briefly, the cells were lysed for 20 min on ice in RIPA lysis buffer containing a protease inhibitor cocktail and DMSF. The samples were subjected to 12% SDS-PAGE and transferred to nitrocellulose membranes. Blots were probed with primary antibodies anti-Caspase-3 (Abcom, 1:1,000), anti-Cyto c (Abclone, 1:1,000), anti-Tublin (Affinity, 2:1,000), and anti-β-actin (TransGen, 1:2,000) at 4°C overnight with gently shaking. After three times washing with PBS, the horseradish peroxidase (HRP)-conjugated secondary antibodies were added. Antigen-antibody complexes were visualized by enhanced chemiluminescence. Enhanced ECL TM prime detection reagent (30 (link)) was used to visualize antigen-antibody complexes, and the density was quantified by ImageJ.
Anti cyto c
Manufactured by Abclone
Anti-Cyto c is a laboratory reagent used to detect the presence of cytochrome c, a key protein involved in the mitochondrial electron transport chain and apoptosis signaling. It can be used in various analytical techniques, such as Western blotting, ELISA, and immunohistochemistry, to identify and quantify cytochrome c levels in biological samples.
Automatically generated - may contain errors
2 protocols using anti cyto c
1
Immunoblot Analysis of Apoptosis Markers
2
Apoptosis Pathway Protein Detection
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Cells were lysed for 20 minutes on ice in RIPA lysis buffer containing a protease inhibitor cocktail and DMSF. The samples were subjected to 12% SDS-PAGE and transferred to nitrocellulose membranes. Blots were probed with primary antibodies anti-Caspase3 (Abcom, 1:1000), anti-Cyto c (Abclone, 1:1000) anti-Tubulin (A nity, 1:1000) anti-β-actin (TransGen, 1:2000) at 4°C overnight with gently shaking. After three times washing with PBS, the horseradish peroxidase (HRP)-conjugated secondary antibodies were added. Antigen antibody complexes were visualized by enhanced chemiluminescence. Enhanced ECL TM prime detection reagent (GE) used to visualize antigen-antibody complexes, the density quanti ed by Image J.
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