Key features and details
- Mouse monoclonal [F5D] to Myogenin
- Suitable for: IHC-FoFr, ICC, IHC-P, WB
- Reacts with: Mouse, Rat, Human, Pig
- Isotype: IgG1
Product nameAnti-Myogenin antibody [F5D]
See all Myogenin primary antibodies
DescriptionMouse monoclonal [F5D] to Myogenin
Tested applicationsSuitable for: IHC-FoFr, ICC, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Rat, Human, Pig
Recombinant full length protein corresponding to Rat Myogenin aa 1-250.
Database link: P20428
- IHC-P: Human Rhabdomyosarcoma; ICC: 2 days differentiated C2C12 cells, ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612); WB: C2C12 cells post day 2 differentiation into myotubes.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact firstname.lastname@example.org.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
PurityProtein G purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab1835 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||Use at an assay dependent concentration.|
|ICC||Use a concentration of 1 - 5 µg/ml.|
|IHC-P||Use at an assay dependent concentration.|
|WB||1/250 - 1/500. Detects a band of approximately 40 kDa (predicted molecular weight: 25 kDa).|
FunctionInvolved in muscle differentiation (myogenic factor). Induces fibroblasts to differentiate into myoblasts. Probable sequence specific DNA-binding protein.
Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
- Information by UniProt
- bHLHc3 antibody
- cb553 antibody
- Class C basic helix-loop-helix protein 3 antibody
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 :
Lane 1 : Differentiated C2C12 whole cell lysate - Day 0 control
Lane 2 : Differentiated C2C12 whole cell lysate - Day 1
Lane 3 : Differentiated C2C12 whole cell lysate - Day 2
Lane 4 : Differentiated C2C12 whole cell lysate - Day 3
Lane 5 : Differentiated C2C12 whole cell lysate - Day 4
Lane 6 : Differentiated C2C12 whole cell lysate - Day 5
Lane 7 : Differentiated C2C12 whole cell lysate - Day 6
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 34 kDa why is the actual band size different from the predicted?
C2C12 cells were differentiated into myotubes as previously described in PMID:26563778.
Lanes 1-7: Merged signal (red and green). Green - ab1835 observed at 30kDa. Red - loading control ab181602 observed at 37kDa.
This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab1835 and ab181602 (Rabbit anti GAPDH loading control), were incubated overnight at 4°C at a 1 in 400 dilution and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunofluorescence staining of Myogenin using ab1835 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 3 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 ab1835 at 1 µg/mL and ab6046, rabbit polyclonal to beta Tubulin, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150088, Goat Anti-Rabbit 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 ab1835 gave comparable results using MeOH fixation (100%, 5 min).
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.
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.
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 has been referenced in 48 publications.
- Zhou H et al. Chromatin accessibility is associated with the changed expression of miRNAs that target members of the Hippo pathway during myoblast differentiation. Cell Death Dis 11:148 (2020). PubMed: 32094347
- Li J et al. Long Non-Coding RNA H19 Promotes Porcine Satellite Cell Differentiation by Interacting with TDP43. Genes (Basel) 11:N/A (2020). PubMed: 32121115
- Nakagawa T et al. Administration of Slow-Release Synthetic Prostacyclin Agonist Promoted Angiogenesis and Skeletal Muscle Regeneration for Limb Ischemia. Mol Ther Methods Clin Dev 18:119-130 (2020). PubMed: 32637444
- He J et al. Transcriptome Characterization of Repressed Embryonic Myogenesis Due to Maternal Calorie Restriction. Front Cell Dev Biol 8:527 (2020). PubMed: 32671071
- Combes AN et al. Single-cell analysis reveals congruence between kidney organoids and human fetal kidney. Genome Med 11:3 (2019). PubMed: 30674341