Recombinant Anti-mtTFA antibody [EPR12285] - Mitochondrial Marker (ab176558)

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST

Immunofluorescence staining of MCF7 cells with purified ab176558 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab176558 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

ab176558 (purified) at 1/40 immunoprecipitating mtTFA in 10 μg HEK293 (Lanes 1 and 2, observed at 24 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue, labeling mtTFA using unpurified ab176558 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human prostate tissue, labeling mtTFA using unpurified ab176558 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunofluorescence analysis of MCF7 cells, labeling mtTFA using unpurified ab176558 at a 1/100 dilution.

Western blot analysis on Immunoprecipitation pellet from either 1) 293T cell lysate, or 2) 1xPBS (negative control), showing mtTFA, using unpurified ab176558 at 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.