Recombinant HRP Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab195024)

ab195024 was shown to recognize LRRK2 in wild-type cells as signal was lost at the expected MW in LRRK2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LRRK2 knockout samples were subjected to SDS-PAGE. Ab195024 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor^® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

IHC image of LRRK2 staining in a section of formalin-fixed paraffin-embedded normal human cerebellum. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab195024 at 1/500 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195024 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.