ab40657 staining IGF1 in murine prostate tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed with paraformaldehyde and an antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% serum for 20 minutes at 25°C and then incubated with ab40657 at a 1/1000 dilution for 18 hours at 4°C. The secondary used was a biotin conjugated horse polyclonal used at a 1/50 dilution.See Abreview
ab40657 stained Hek293 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 ab40657 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.
Immunohistochemical analysis of rat colon tissue, staining IGF1 with ab40657.Tissue was fixed with formaldehyde and blocked with 1% blocking solution for 15 minutes at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/400 in BSA in TBS) for 1 hour. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody. See Abreview