Anti-Myogenin antibody [F5D] (ab1835)

ab1835 staining Myogenin in undifferentiated C2C12 cells (top panel) and 2 days differentiated C2C12 cells (bottom panel).

The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 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 with ab1835 at 1 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution overnight at 4ºC. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

IHC image of Myogenin staining in a section of formalin-fixed paraffin-embedded normal Human Rhabdomyosarcoma* performed on a Leica BOND™ system using the standard Protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab1835, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab1835 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

ab1835 at 1/50 dilution staining pig differentiated skeletal myoblast cells by ICC/IF. The cells were paraformaldehyde fixed, permeabilized with Triton-X100 and incubated with the antibody overnight at 4C. An Alexa Fluor 488 conjugated goat anti-mouse antibody was used as the secondary. The image was captured using a confocal laser scanning microscope with an additional Differential Interference Contrast (DIC) mode. The image shows DAPI nuclear counterstain (blue, upper left panel), Myogenin staining (green, upper right panel), DIC (phase) image (lower left panel) and overlay (lower right panel).

Myogenin is not expressed in proliferating myoblasts, but rather in mononucleated differentiating myoblasts (See abreview for more detail).

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