The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.5 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
Use a concentration of 1 - 2 µg/ml.
Use at an assay dependent concentration. PubMed: 21490313
Use a concentration of 1 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
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.
ab14730 staining ATPB in cultured human fibroblasts. Cells were fixed, permeabilized and then labelled with ab14730 followed by an AlexaFluor® 488-conjugated Goat anti-Mouse IgG1-specific secondary antibody (2 µg/ml)
Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
All lanes : Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
Lane 1 : isolated mitochondria from human heart at 5 µg Lane 2 : isolated mitochondria from bovine heart at 1 µg Lane 3 : isolated mitochondria from rat heart at 10 µg Lane 4 : isolated mitochondria from mouse heart at 10 µg
Predicted band size: 52 kDa Observed band size: 52 kDa
Immunohistochemistry (Frozen sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)This image is courtesy of an anonymous Abreview
ab14730 staining ATPB in rat liver tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 10% goat serum) for 18 hours at 4°C. An Alexa Fluor® 555-conjugated goat anti-mouse polyclonal (1/500) was used as the secondary antibody.
ab14730 (2µg/ml) staining ATPB in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic and mitochondrial staining of epithelium. Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.