All lanes : Anti-Cyclophilin A antibody (ab42408) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : Lymph node (Human) Tissue Lysate - adult normal tissue (ab29871)Lane 4 : Thymus (Human) Tissue Lysate - adult normal tissue (ab30146)Lane 5 : Lung (Rat) Tissue LysateLysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
Anti-Cyclophilin A antibody (ab42408) at 1/1000 dilution + Human Cyclophilin A full length protein (ab96022) at 0.1 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 1 minute
ICC/IF image of ab42408 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42408, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
ab42408 staining Cyclophilin A in human heart tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval and then blocked (5 minutes of peroxidase block and 10 minutes of protein block) for 15 minutes at 20°C. The primary antibody was used undiluted and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used, undiluted as secondary. See Abreview