Anti-Myogenin 抗体 [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).

All lanes : Anti-Myogenin antibody [F5D] (ab1835) at 1/250 dilution

Lane 1 : Skeletal Muscle (Mouse) Tissue Lysate
Lane 2 : Skeletal Muscle (Rat) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 25 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?


Exposure time: 12 minutes

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

Adult mouse muscle section stained with  ab1835. The animals were perfused with 4% PFA. The sections were incubated in 5% normal donkey serum in 0.1% PBS- and triton X100 for 1h to permeabilise the cells and block non-specific protein-protein interactions. The sections were then incubated with the antibody (ab1835, 1µg/ml) overnight at +4°C. The secondary antibody Alexa Fluor® 568 donkey anti-mouse IgG (H+L) (red) was used at a 1/1000 dilution for 1h.

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).

See Abreview