53-5643-80

antibody from Invitrogen Antibodies

Targeting: MYOG bHLHc3, MYF4

Provider product page for 53-5643-80

Immunocytochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [4]
Comments [0]
Validations
Immunocytochemistry [2]
Other assay [5]

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Product number: 53-5643-80 - Provider product page
Provider: Invitrogen Antibodies
Product name: Myogenin Monoclonal Antibody (F5D), Alexa Fluor™ 488, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: This F5D monoclonal antibody reacts with human, mouse, dog, and rat myogenin, a 34-kDa transcription factor. Expressed in skeletal and heart muscle, myogenin is a member of the MyoD family of basic-helix-loop-helix proteins, which also includes MyoD, Myf5, and MRF4. This transcription factor interacts with other helix-loop-helix proteins, which may or may not be muscle-specific. Myogenin plays a significant role in myogenic differentiation, even directing nonmuscle cells to the myogenic lineage. Transforming growth factor-beta (TGFb) and bone morphogenetic protein-2 (BMP2) inhibit myogenin transcriptional activity. Predominantly residing within the nucleus, the subcellular localization of myogenin has been shown to be dependent on differentiation status and cell density. For instance, trafficking of myogenin between the nucleus and cytoplasm has been reported during skeletal muscle differentiation to mediate transcription control. Applications Reported: This F5D antibody has been reported for use in immunocytochemistry. Applications Tested: This F5D antibody has been tested by immunocytochemistry of the C2C12 cell line. This can be used at less than or equal to 5-20 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Excitation: 488 nm; Emission: 519 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human, Mouse, Rat, Canine
Host: Mouse
Conjugate: Green dye
Isotype: IgG
Antibody clone number: F5D
Vial size: 25 µg
Concentration: 0.5 mg/mL
Storage: 4° C, store in dark, DO NOT FREEZE!

MYOG protein structure - 53-5643-80 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 The effect of 20 muM PUFA on C2C12 myotubes formation. ( a ) Immunofluorescence staining using Anti-Myogenin Alexa Fluor (r) 488 and Hoechst 33342 staining on C2C12 myotubes at day 2 (d2) of the treatment. Scale bar for live cells is 100 mum, Scale bar for anti-myogenin Alexa488, Hoechst 33342 staining and merged is 200muM. ( b ) Quantification of differentiation index and fusion index are shown. The mRNA level of myogenesis markers at three time points (d2 (2 days treatment), d4 (2 days treatment plus 2 days recovery), d8 (2 days treatment plus 6 days recovery)) was showed. ( c ) At d2, Myf5 expression level was induced in both AA20 and DHA20 treatments. MyoG expression level was significantly decreased in DHA20 treatment. MyoD1 expression level was decreased in AA20 treatment. The expression of Myf6 significantly decreased at d4 in DHA20, and significantly decreased in both AA20 and DHA20 at d8. ( d ) The expression levels of Myh1 significantly decreased at d2, d4 and d8 in DHA20 and in AA20 at d8. The expression levels of Myh2 significantly increased in d4 and d8 in AA20 and significantly decreased in DHA20 at d8. The expression levels of Myh4 significantly decreased at d4 in DHA20 and in both AA20 and DHA20 at d8. All values are LS means +- SEM, + indicate p < 0.1 and * indicates p < 0.05 treatment vs. control group BSA20.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 The effect of 50 muM PUFA on C2C12 myotubes formation. ( a ) Immunofluorescence staining using Anti-Myogenin Alexa Fluor (r) 488 and Hoechst 33342 staining on C2C12 myotubes at day 2 (d2) of the treatment. Scale bar for live cells is 100 mum, Scale bar for anti-myogenin Alexa488, Hoechst 33342 staining and merged is 200 muM. ( b ) Quantification of differentiation index and fusion index are shown. The mRNA level of myogenesis markers at three time points (d2 (2 days treatment), d4 (2 days treatment plus 2 days recovery), d8 (2 days treatment plus 6 days recovery)) was shown. ( c ) At d2, Myf5 expression level was induced in both AA50 and DHA50 treatment. MyoG expression level was significantly decreased in both AA50 and DHA50 treatment. MyoD1 expression level was significantly decreased in AA50 and DHA50 treatments. The expression level of Myf6 was significantly increased in DHA50 at d2 and decreased at d4, but significantly decreased in both AA50 and DHA50 at d8. ( d ) The expression level of Myh1 was significantly decreased in d2, d4 and d8 in DHA50, also significantly decreased in AA50 at d4. The expression level of Myh2 was significantly increased in AA50 and significantly decreased in DHA50 at d8. The expression levels of Myh4 significantly decreased at d4 and d8 in both AA50 and DHA50. All values are LS means +- SEM, + indicate p < 0.1 and * indicates p < 0.05 treatment vs. control group BSA50.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 2 Metformin inhibits myogenic differentiation of C2C12 cells in a dose-dependent manner. (a) Immunofluorescence microscopy for the expression of the myogenic markers Myosin Heavy Chain (MHC) and MyoD. C2C12 induced to differentiate by serum deprivation in differentiation medium (DM) and treated for 3 days with different doses of metformin (100muM, 500muM, 2mM, 5mM and 10mM) were analyzed by immunofluorescence technique. The percentage of the cells positive for MyoD and the fusion index were determined using the Cell Profiler cell image analysis software. The fusion index was calculated as the % of the nuclei inside myotubes compared to the total number of nuclei. Only myotubes with at least three nuclei inside a continuous cell membrane were considered. Statistical significance was evaluated by the Student's t-test (*p
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 IPA-3 treatment reduces p38 phosphorylation during Mabs and C2C12 differentiation. Representative western blot revealing total p38, p-p38, and myogenin expression in crude protein extracts from IPA-3 treated or control Mabs (A) and C2C12 (D) during 5 days of myogenic differentiation. beta-Tubulin, vinculin, or actin are used as loading controls. The bar graphs illustrate the ratio between p-p38 and total p38 signals during Mabs (B) or C2C12 (E) differentiation, as determined by the densitometric quantitation of western blots such as the one in panel A and D. The bar graphs represent the densitometric quantitation of myogenin expression for Mabs (C) and C2C12 (F) . The values are mean of at least three independent experiments + SEM. Statistical significance was evaluated by the unpaired t -test, * P
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Budesonide treatment promotes terminal differentiation of mdx satellite cells through the activation of GCr. ( a ) SCs were isolated from muscles of mdx mice as CD45-/CD31-/ITGA7 + cells and plated in sGM. 48 hours after plating, cells were treated with three concentrations of budesonide (0.1, 1 and 5 muM) or TSA (20 nm) for 5 additional days. Myogenic differentiation was assessed by immunostaining with antibodies against myogenin ( b ) and MyHC ( c ) as late muscle-specific differentiation markers. Nuclei were counterstained with Hoechst 33342. ( d ) Column chart showing the percentages of myogenin positive cells in the experiment in panel b. ( e ) Bar plot reporting the number of nuclei per field for the experiments in b and c. ( f,g ) Bar plots showing the fusion index and myotube diameter for the experiment in c. The values are mean of three independent experiments +- SEM (n = 3). Statistical significance was evaluated using one-way ANOVA (*p
Conjugate: Green dye