All lanes : Anti-SPHK1 antibody (ab16491) at 1 µg/mlLane 1 : Human Skeletal muscle lysateLane 2 : Human Skeletal muscle lysate with Human SPHK1 peptide (ab16634) at 1 µg/mlLysates/proteins at 20 µg/ml per lane.SecondaryAlexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution

ab16491 staining human smooth muscle, mouse lung, and rat heart tissue sections by IHC-P.  Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking in sequential peroxidase and protein block (prediluted) for 20 minutes at 20°C.  The primary antibody was diluted 1/100 and incubated with the sample for 45 minutes at 20°C.  A HRP-conjugated goat anti-rabbit antibody was used as the secondary.See Abreview

ICC/IF image of ab16491 stained HepG2 cells. The cells were 4% PFA 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 (ab16491, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit 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.