The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Recommended starting point for ab1835 incubation is 30min RT.
1/250. Detects a band of approximately 40 kDa (predicted molecular weight: 25 kDa).
Abcam recommends using BSA as the blocking agent.
Use a concentration of 1 µg/ml.
Involved in muscle differentiation (myogenic factor). Induces fibroblasts to differentiate into myoblasts. Probable sequence specific DNA-binding protein.
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
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Myogenin antibody [F5D] (ab1835)This image is courtesy of Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom
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.
Immunocytochemistry/ Immunofluorescence - Anti-Myogenin antibody [F5D] (ab1835)This image is courtesy of an Abreview submitted by Dr Mal Niladri
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).