Anti-Desmin antibody - Cytoskeleton Marker (ab15200)

Immunohistochemical analysis of paraffin-embedded on human skeletal muscle labeling Desmin with ab15200 at 1/200 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin.

Immunofluorescence staining of Desmin using ab15200 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 10 days post induction.

The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab15200 at 0.1 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.

The antibody ab15200 gave comparable results using MeOH fixation (100%, 5 min).

Immunocytochemistry/Immunofluorescence analysis of C2C12 (mouse muscle) cells labelling Desmin with ab15200 at 1/500. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

ab15200 staining rat differentiated skeletal muscle cells by ICC/IF.

Cultured differentiated skeletal muscle cells were harvested from Lewis rats, 2% paraformaldehyde treated for 15 min to fix the cells, and then permealized with Triton-X100 (0.1%) for 10 min. The cells were then incubated with ab15200 at 1/200 overnight at 4°C. The image was taken with a confocal laser scanning microscope and shows desmin expressing skeletal myocytes (green-cytoplasmic localization). Note that desmin forms short thickened filamentous structures in the cell cytoplasm, along with prominent spot-like cytoplasmic aggregates that are composed of densely packed filaments.

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Immunocytochemistry/Immunofluorescence analysis of RMS13 (human rhabdomyosarcoma) cells labelling Desmin with ab15200 at 1/500. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells (negative cell line) showing weak staining of Desmin with ab15200 at 1/500. Cells were fixed with 100% methanol. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

ab15200 staining Desmin in rat Smooth muscle cells from mesenteric artery by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 4% paraformaldehyde in PSS for 4 min at 4°C and permeabilized with 0.3% Triton x100 before blocking with 2% BSA was done for 30 minutes at 20°C. Samples were incubated with primary antibody (1/300 in PSS with 2%BSA and 0.3% Triton X-100) for 14 hours at 4°C. An MFP 555-conjugated donkey polyclonal to rabbit IgG was used as secondary antibody at 1/400 dilution.

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Immunohistochemical analysis of paraffin-embedded on human cardiac muscle labeling Desmin with ab15200 at 1/200 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded on rat cardiac muscle labeling Desmin with ab15200 at 1/200 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin.

ab15200 staining Desmin in Mouse skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature; antigen retrieval was enzymatic. Samples were incubated with primary antibody (1/800 in blocking buffer) for 16 hours at 4°C. An undiluted HRP-conjugated Horse anti-rabbit IgG polyclonal was used as the secondary antibody.

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Immunohistochemical analysis of PFA-fixed paraffin-embedded murine nephric tissue, labelling Desmin with ab15200 at a dilution of 1/250 incubated for 18 hours at 4°C. Heat mediated antigen retrival was with citrate buffer. Blocking was with Dako serum-free protein block at 100% incubated for 1 hour at 23°C. Secondary was a donkey anti-rabbit polyclonal HRP conjugate at 1/200.

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