E. coli- derived recombinant human MyoD, Glu77-Gly215 (P15172). Species Sequence Homology: mouse, rat (96%).
MyoD (myoblast determination protein 1), also called Myf-3 (myogenic factor 3) is an approximately 44kD nuclear protein in the MyoD family of muscle-specific bHLH transcription factors. MyoD family members heterodimerize with E proteins and cooperate with MEF2 family transcription factors to regulate expression of skeletal muscle-specific genes. MyoD is essential for skeletal muscle differentiation. Acetylation at lysines 99, 102 and 104 further regulates its activity. Applications: Suitable for use in Immunocytochemistry. Other applications not tested. Recommended Dilution: Immunocytochemistry: 10ug/ml detects MyoD in C2C12 mouse muscle myoblast cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at RT and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT. After blocking, cells were incubated with diluted M9780-05A overnight at 4C followed by fluorochrome-coupled anti-mouse IgG at RT in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20C. Stable for 12 months at -20C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
