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Semi purified mitochondrial preparation.
Our Abpromise guarantee covers the use of ab3298 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FrFl||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. Used at a dilution of 1/50 for 2 hours (see Abreview for further information).|
|ICC||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 - 2 µg/ml. PubMed: 18167556Samples require boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10 - 20 min followed by cooling at RT for 20 min. This antibody can be used to stain themitochondria in cell / tissue preparations and can be used as a marker of the mitochondria in subcellular fractions.|
|IHC-Fr||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 - 2 µg/ml. This antibody detects a band of 60 kDa, which corresponds to the predicted molecular weight of the non glycosylated protein component of mitochondria.|
|Flow Cyt||Use 1µg for 106 cells.|
ab3298 staining Mitochondria in Human Fibroblast by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldeshyde, permeabilized with 0.1% Triton X-100 and blocked with 1% milk for 15 minutes. Samples were incubated with primary antibody (1/50 in 1% milk) for 1 hour at 21°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.
ab3298 staining Mitochondria (red) in A549 Human alveolar basal epithelial cells by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 1 hour at 25°C; permeabilized with 0.3% Triton X-100, 0.1% Saponin. Samples were incubated with primary antibody (1/500 in 1% BSA, 0.3% Triton X-100, 0.1% Saponin) for 16 hours at 4°C. A DyLight 649-conjugated Goat anti-mouse IgG1 polyclonal (1/300) was used as the secondary antibody. DAPI (blue), actin stained with fluorescent phalloidin (green)
Mitochondria were detected by Immunofluorescence in U373 human glioblastoma cells using Mitochondrial Marker antibody (ab3298).
ab3298 at 0.2µg/ml staining Mitochondria-Mitochondrial Marker from human cardiac muscle sections by Immunohistochemistry (Paraffin-embedded sections). The antibody was incubated with the tissue for 2 hours and then detected with an Alexa-Fluor® 488 conjugated anti-mouse IgG secondary antibody.
This image is courtesy of an Abreview submitted by an anonymous researcher on 12 December 2005.
ab3298 staining HEK 293 cells by ICC/IF. Following paraformaldehyde fixation and permeabilisation with 0.5%TX100 the cells were blocked with 5% serum (for 20 hours at 25°C). The primary antibody was diluted 1/100 in PBS, 2% goat serum, 0.1% TX100 and incubated for 1 hour at 25°C. An Alexa Fluor® 546 conjugated goat anti-mouse was used as the secondary.
Mitochondria are labeled with red fluorescence, and the nuclei were counterstained with DAPI.
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