The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 25223649
Use a concentration of 0.5 - 1 µg/ml.
Perform heat-induced antigen-retrieval with 100mM Tris, 5% urea, pH 9.5 at 95°C for 10 minutes immediately before permeabilization.
Use a concentration of 0.5 µg/ml. Predicted molecular weight: 23 kDa.
Use a concentration of 1 µg/ml. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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. Part of the complex F(0) domain and the peripheric stalk, which acts as a stator to hold the catalytic alpha(3)beta(3) subcomplex and subunit a/ATP6 static relative to the rotary elements.
Human colon tissue fixed with 4% PFA and antibody detection with ab110276 using the ABC system. ab110276 diluted 1/500 and incubated for 1 hour. Sections were incubated in peroxidase-conjugated rabbit anti-mouse immunoglobulins (diluted 1/100 in 4% BSA in PBST) for 1 hour at room temp.
Mitochondrial localization of ATP5O. Cultured Human embryonic lung-derived fibroblasts (strain MRC5) were fixed, treated for heat-induced antigen retrieval, permeabilized and then labeled with ab110276 (0.5 µg/ml) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG1 isotype specific secondary antibody (2 µg/ml).
HL-60 cells were stained with 1 µg/mL ab110276 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
Western blot - Anti-ATP5O antibody [4C11C10D12] (ab110276)
All lanes : Anti-ATP5O antibody [4C11C10D12] (ab110276) at 1 µg/ml
Lane 1 : HDFn (human) cell lysates at 20 µg Lane 2 : COS7 (monkey) cell lysates at 20 µg Lane 3 : H4IIE (rat) cell lysates at 20 µg Lane 4 : MEF (mouse) cell lysates at 20 µg Lane 5 : bovine heart mitochondria lysates at 5 µg