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Flow cytometric analysis of myogenin expression in human rhabdomyosarcoma cells. Human RH-30 rhabdomyosarcoma cells (DSMZ, Cat. No. ACC 489) were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527), fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885). The cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Myogenin antibody (Cat. No. 563120; solid line histogram) at matched concentrations. The histograms were derived from gated events with the light scattering characteristics of intact RH-30 cells. Flow cytometry was performed using a BD LSRFortessa™ II Flow Cytometry System. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization of fixed cells. However, a small increase in non-specific background staining may occur in some negative cell lines.
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BD Pharmingen™ PE Mouse anti-Myogenin
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Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
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Myogenin is a member of the MyoD family of myogenic basic helix-loop-helix (bHLH) transcription factors that also includes MyoD, Myf-5, and MRF4 (also known as herculin or Myf-6). MyoD family members are expressed exclusively in skeletal muscle and play a key role in activating myogenesis by binding to enhancer sequences of muscle-specific genes. The regulatory domain of MyoD is approximately 70 amino acids in length and includes both a basic DNA binding motif and a bHLH dimerization motif. MyoD family members share about 80% amino acid homology in their bHLH motifs. Transfection of myogenin and other family members into a variety of non-muscle cells has been shown to either convert these cells to myogenic cells or to transcriptionally activate a set of otherwise unexpressed muscle-specific genes. In addition to activating muscle-specific genes, members of the MyoD family members activate their own transcription and transactivate the transcription of other MyoD family members. For example, transfection of myogenin into 10T1/2 cells or Swiss 3T3 cells results in the activation of the endogenous myogenin gene as well as transactivation of MyoD. Likewise, the transfection of MyoD into these cells results in the activation of MyoD as well as the transactivation of myogenin. Each member of the MyoD family has distinct roles in muscle development; myogenin plays a key role in muscle maturation. Myogenin migrates at a molecular weight of ~34 kDa by SDS-PAGE.
The F5D monoclonal antibody recognizes human, mouse, rat and cat myogenin. Mapping studies suggest that F5D reacts with an epitope between amino acids 138 and 158 of myogenin.

Development References (5)
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Dias P, Dilling M, Houghton P. The molecular basis of skeletal muscle differentiation.. Semin Diagn Pathol. 1994; 11(1):3-14. (Biology). View Reference
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Gang EJ, Jeong JA, Hong SH, et al. Skeletal myogenic differentiation of mesenchymal stem cells isolated from human umbilical cord blood. Stem Cells. 2004; 22(4):617-624. (Clone-specific: Flow cytometry). View Reference
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Meadows E, Cho JH, Flynn JM, Klein WH. Myogenin regulates a distinct genetic program in adult muscle stem cells. Dev Biol. 2008; 322(2):406-414. (Biology). View Reference
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Ryan T, Liu J, Chu A, Wang L, Blais A, Skerjanc IS. Retinoic acid enhances skeletal myogenesis in human embryonic stem cells by expanding the premyogenic progenitor population. Stem Cell Rev. 2012; 8(2):482-493. (Biology). View Reference
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Wang NP, Marx J, McNutt MA, Rutledge JC, Gown AM. Expression of myogenic regulatory proteins (myogenin and MyoD1) in small blue round cell tumors of childhood. Am J Pathol. 1994; 147(6):1799-1810. (Clone-specific: Immunohistochemistry). View Reference
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