The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/5000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
Mitochondrial membrane ATP synthase (F(1)F(0) ATP synthase or Complex V) produces ATP from ADP in the presence of a proton gradient across the membrane which is generated by electron transport complexes of the respiratory chain. F-type ATPases consist of two structural domains, F(1) - containing the extramembraneous catalytic core, and F(0) - containing the membrane proton channel, linked together by a central stalk and a peripheral stalk. During catalysis, ATP synthesis in the catalytic domain of F(1) is coupled via a rotary mechanism of the central stalk subunits to proton translocation. Subunits alpha and beta form the catalytic core in F(1). Rotation of the central stalk against the surrounding alpha(3)beta(3) subunits leads to hydrolysis of ATP in three separate catalytic sites on the beta subunits.
Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (HRP) (ab197905)
All lanes : Anti-ATPB antibody [3D5] - Mitochondrial Marker (HRP) (ab197905) at 1/5000 dilution
Lane 1 : Human heart tissue lysate - mitochondrial extract (ab110337) at 5 µg Lane 2 : HeLa whole cell lysate (ab150035) at 10 µg Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 52 kDa Observed band size : 52 kDa
Exposure time : 2 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab197905 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.