Recombinant Anti-Desmin antibody [Y66] - Cytoskeleton Marker (ab32362)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue labelling Desmin with purified ab32362 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Immunofluorescence staining of Desmin using ab32362 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 10 days post induction.

The cells were fixed with 100% MeOH (5 min) 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 ab32362 at 0.02 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 ab32362 gave comparable results using 4% formaldehyde fixation (10 min).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

Unpurified ab32362 staining Desmin (green) in Human skeletal muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methacarn and blocked with 10% serum for 20 minutes at 22°C. Samples were incubated with primary antibody (1/150) for 12 hours. An Alexa Fluor^® 488-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Blue - DAPI-nuclei. Red - WGA. 40X objective.

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Immunocytochemistry/Immunofluorescence analysis of C2C12 (Mouse myoblasts myoblast) cells labeling Desmin with ab32362 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. 

ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 was used as counterstain antibody.

Confocal image showing cytoplasmic staining on C2C12 cell line.

Intracellular Flow Cytometry analysis of C2C12 cells labelling Desmin with purified ab32362 at 1/70 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies. 

Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling Desmin with purified ab32362 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control: primary antibody (1/50) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Immunofluorescent analysis of Human mitochondria injected rabbit hearts sections stained for Desmin (Green) using ab32362. MTCO2, the human-specific mitochondrial marker was stained in red, and the nuclei was stained using the DNA stain DAPI (blue).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

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Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human uterus tissue labelling Desmin with unpurified ab32362.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human urinary bladder tissue labelling Desmin with unpurified ab32362.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human skeletal muscle tissue labelling Desmin with unpurified ab32362.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human brain tissue. Unpurified ab32362 shows negative staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human tonsil tissue. Unpurified ab32362 shows negative staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Unpurified ab32362 staining Desmin (green) in Mouse aorta smooth muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formalin and blocked with 10% serum for 20 minutes at 22°C. Samples were incubated with primary antibody (1/150) for 1 hour at 22°C. An Alexa Fluor^® 488-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Blue - nuclei.

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