製品の概要

  • 製品名

    Anti-Myogenin antibody [F5D]
    Myogenin 一次抗体 製品一覧
  • 製品の詳細

    Mouse monoclonal [F5D] to Myogenin
  • 由来種

    Mouse
  • アプリケーション

    適用あり: Flow Cyt, IHC-P, ICC/IF, WB, IHC-FoFrmore details
  • 種交差性

    交差種: Mouse, Rat, Human, Pig
  • 免疫原

    Recombinant full length protein corresponding to Rat Myogenin aa 1-250.

  • ポジティブ・コントロール

    • IHC-P: Human Rhabdomyosarcoma ICC/IF: 2 days differentiated C2C12 cells
  • 特記事項

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

     

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Flow Cyt 1/50.
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
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 a concentration of 1 µg/ml.

ターゲット情報

画像

  • 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.

  • 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

参考文献

This product has been referenced in:

  • Combes AN  et al. Single-cell analysis reveals congruence between kidney organoids and human fetal kidney. Genome Med 11:3 (2019). Read more (PubMed: 30674341) »
  • McClure MJ  et al. Integrin-a7 signaling regulates connexin 43, M-cadherin, and myoblast fusion. Am J Physiol Cell Physiol 316:C876-C887 (2019). Read more (PubMed: 30892939) »
See all 41 Publications for this product

レビューと Q&A

1-10 of 10 Abreviews or Q&A

Application
Flow Cytometry
Sample
Pig Cell (Muscle)
Permeabilization
Yes - 0.1% Triton X-100 / PBS
Gating Strategy
Negative control (cells incubated with normal serum instead of primary antibody)
Specification
Muscle
Preparation
Cell harvesting/tissue preparation method: Cells detached with HyQtase, 10% FBS / DPBS added, centrifugation of cells, fixation in 4% PFA (30 min, 4°C) wash with PBS 3 times, permeabilization 0.1% Triton X-100 / PBS (10 min, RT) blocking 2% rabbit serum / 0.1% Triton X-100 / PBS (60 min, RT) incubation with antibody
Sample buffer: 2% Rabbit Serum / 0.1% Triton X-100 / PBS
Fixation
Paraformaldehyde

Abcam user community

Verified customer

投稿 Oct 26 2016

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

Abcam user community

Verified customer

投稿 Jan 05 2011

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Muscles)
Specification
Muscles
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% TritonX100

Dr. Ruma Raha-Chowdhury

Verified customer

投稿 May 19 2010

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

Dr. Ruma Raha-Chowdhury

Verified customer

投稿 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

Dr. Henrike Sell

Verified customer

投稿 Feb 02 2010

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%

Dr. Mal Niladri

Verified customer

投稿 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%

Dr. Mal Niladri

Verified customer

投稿 Oct 09 2006

Answer

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 rhabdomyosarcomas. You can refer to the following publications for more information: Wright WE, et al. Dev Genet 19: 131-138 (1996). Wright WE, et al. Cell 1989; 56:607. Dr. Wright is the original developer of this clone.

Read More

Answer

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 also positively stains the nuclei of myoblasts from human fetal limbs. This antibody would be a good choice for this customer's purposes and it is also suitable for labeling cryostat sections.

Read More

Answer

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 dilution to recommend-the optimal dilutions / concentrations should be determined by the end user. If you have any more questions, please contact us again.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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