Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testicular seminoma tissue labelling SRPK1 with ab90527 at 1/200 (1µg/ml). Detection: DAB.
ICC/IF image of ab90527 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab90527 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in methanol fixed (100%, 5min) HepG2, Hek293, MCF-7 cells at 1ug/ml, and also in formaldehyde fixed (4%, 10min) HeLa, Hek293, HepG2, MCF-7 cells at 1ug/ml.
All lanes : Anti-SRPK1 antibody (ab90527) at 0.04 µg/mlLane 1 : HeLa whole cell lysate at 50 µgLane 2 : HeLa whole cell lysate at 15 µgLane 3 : HeLa whole cell lysate at 5 µgLane 4 : Whole cell lysate from 293T cells at 50 µgLane 5 : Whole cell lysate from NIH 3T3 cells at 50 µgdeveloped using the ECL technique
ab90527 at 1 µg/ml detecting SRPK1 following immunoprecipitation. Detection by ECL with a 3 second exposure. Lane 1; IP with ab90527 Lane 2; IP with an antibody to a downstream epitope of SRPK1 Lane 3; IP with control IgG All antibodies used at at 3 µg/mg lysate for immunoprecipitation. 1 mg of lysate was used for IP and 20% of IP loaded.