Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma (left) and mouse teratoma tissues labelling USP13 with ab99421 at 1/5000 (0.2µg/ml) and 1/1000 (1µg/ml). Detection: DAB.
ICC/IF image of ab99421 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 ab99421 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.
Western blot - USP13 antibody (ab99421)
All lanes : Anti-USP13 antibody (ab99421) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 97 kDa
Exposure time: 3 minutes
Immunoprecipitation - USP13 antibody (ab99421)
ab99421 at 1 µg/ml detecting USP13 in HeLa whole cell lysate by western blot analysis following immunoprecipitation. Detection utilised ECL with a 10 second exposure.
For immunoprecipitation, ab99421 was used at at 3 µg/mg lysate; 1 mg of lysate was used for IP and 20% of IP was loaded.
Lane 1; IP using ab99421
Lane 2; IP using ab99423, a rabbit anti-USP13 antibody which recognizes a downstream epitope.
Lane 3; IP using control IgG.