ICC/IF image of ab74020 stained PANC-1 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab74020 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-CDKAL1 antibody (ab74020) at 1 µg/mlLane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : Raji (Human Burkitt's lymphoma cell line) Whole Cell LysateLane 4 : SK N BE (Human neuroblastoma) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
ab74020 staining CDKAL1 in mouse pancreatic tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval and blocking (5 minutes/peroxidase block and 10 minutes/protein block) for 45 minutes at 200C. The primary antibody was diluted 1/250 and incubated with the sample for 45 minutes at 200C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.See Abreview
ab74020 staining CDKAL1 in human pancreatic tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval and blocking (5 minutes/peroxidase block and 10 minutes/protein block) for 15 minutes at 200C. The primary antibody was diluted 1/250 and incubated with the sample for 45 minutes at 200C. A HRP polymer conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.See Abreview