Overview

  • Product nameAnti-Myogenin antibody [F5D]
    See all Myogenin primary antibodies
  • Description
    Mouse monoclonal [F5D] to Myogenin
  • Tested applicationsSuitable for: ICC/IF, Gel supershift assays, IHC-P, IP, WB, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cat, Human, Pig
  • Immunogen

    BALB/ C mice were injected with recombinant protein containing rat myogenin amino acid 30 - 224.

  • Positive control
    • Rhabdomyosarcoma

Properties

Applications

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.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.
Gel supershift assays Use at an assay dependent concentration.
IHC-P 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.
IP Use at an assay dependent concentration.
WB 1/250. Detects a band of approximately 40 kDa (predicted molecular weight: 25 kDa).

Abcam recommends using BSA as the blocking agent.

IHC-FoFr Use at an assay dependent concentration.

Target

Anti-Myogenin antibody [F5D] images

  • 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.
  • 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
    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

  • All lanes : Anti-Myogenin antibody [F5D] (ab1835) at 1/500 dilution

    Lane 1 : mouse muscle lysate
    Lane 2 : mouse muscle lysate
    Lane 3 : mouse muscle lysate
    Lane 4 : mouse muscle lysate
    Lane 5 : mouse muscle lysate
    Lane 6 : mouse muscle lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP conjugated at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 25 kDa
    Observed band size : 25 kDa


    Exposure time : 2 minutes

    Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom

    Non-specific bands detected 37, 64, 74, 95kDa
  • ab1835 - immunohistochemistry

    Formalin fixed paraffin embedded human rhabdomyosarcoma stained with Myogenin using ABC and AEC chromogen.

  • 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

References for Anti-Myogenin antibody [F5D] (ab1835)

This product has been referenced in:
  • Ito A  et al. Induction of functional tissue-engineered skeletal muscle constructs by defined electrical stimulation. Sci Rep 4:4781 (2014). WB ; Mouse . Read more (PubMed: 24759171) »
  • Yang Z  et al. Screening with a novel cell-based assay for TAZ activators identifies a compound that enhances myogenesis in C2C12 cells and facilitates muscle repair in a muscle injury model. Mol Cell Biol 34:1607-21 (2014). WB, ICC/IF ; Mouse . Read more (PubMed: 24550007) »

See all 15 Publications for this product

Product Wall

Application Western blot
Sample Mouse Tissue lysate - whole (skeletal muscle)
Loading amount 30 µg
Specification skeletal muscle
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Jan 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Muscles)
Specification Muscles
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% TritonX100
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Muscle)
Loading amount 20 µg
Specification Muscle
Gel Running Conditions Reduced Denaturing (12% Gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Mar 24 2010

Application Western blot
Sample Human Cell lysate - whole cell (skeletal myotubes)
Loading amount 5 µg
Specification skeletal myotubes
Gel Running Conditions Reduced Denaturing (10 % gel, semi-dry)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Henrike Sell

Verified customer

Submitted Feb 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Pig Cell (Differentiated skeletal myoblast)
Specification Differentiated skeletal myoblast
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%
Username

Dr. Mal Niladri

Verified customer

Submitted Oct 16 2006

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (skeletal myoblast transplanted heart tissue)
Specification skeletal myoblast transplanted heart tissue
Fixative Formaldehyde
Antigen retrieval step Heat mediated
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Username

Dr. Mal Niladri

Verified customer

Submitted Oct 09 2006

This antibody has only been shown to positively stain the nuclei of myoblasts from human fetal tissue but no reactivity has been observed in adult skeletal muscle. We generally use this antibody in-house for detecting myogenin in the nuclei of rhabdom...

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Myogenin is expressed in both normal and tumor cells. Myogenin is expressed in high levels in fetal skeletal muscle, and this antibody has been used to stain human, mouse, and rat cultured myotubes and neonatal mouse, rat, and cat tissue. This clone ...

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Thank you for your enquiry. Ab1835 has been tested for application in Western blotting and shown to cross-react with rat, mouse, and cat (it has not yet been tested in other species). For this antibody in Western blotting, we don't have a specific dilu...

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