Accession # NP_542452; Myoblast determination protein 1, Myogenic differentitaion1,
The myogenic regulatory factors (MRFs) are a family of vertebrate proteins comprise of MyoD, Myf5, Mrf4 and Myog, that are potent transcription factors for muscle genes (1). MyoD, Myf5 and Mrf4 have overlapping but developmentally distinct functions in the specification and differentiation of myoblasts, whereas Myog and Mrf4 activate and maintain the terminal differentiation of muscle (2). MyoD is a muscle specific transcription factor that is essential for vertebrate myogenesis. Certain cell type can be morphologically differentiated into muscle cells by over expression of the bHLH transcription factor MyD1 whereas other cell types such as hepatocytes cannot (2). MyoD1 induces a number of immortalized non muscle cells to e3xpress desmin and several other myofibrillar proteins and caused cells to fuse in to myosacs. The current interest in alteration of cell identity resulted in new approaches to induce pluripotent stem cells that can be transformed into different specialized cell types. The transacription factor MyoD1became an important target for these studies. MyoD1 transform normal dermal fibroblast, chondroblasts, gizzard smooth muscle, and pigmented retinal epithelial cells into elongated postmitotic mononucleated striated myoblasts (3)
The MyoD1 gene encodes a nuclear protein that belongs to a basic helix-loop-helix family of transcription factors and the myogenic factors family. It is involved in muscle growth and regeneration. The transcription of MyoD1 is auto-activated that stabilize commitments to myogenesis. MyoD1 is also known as casual regulator of myogenesis as for other fundamental biological process such as for cell cycle E2F1, glycolysis (HLF), mitochondrial transcription (TFBM2), adoipogenesis (PIAS2), immune function (IRF2) etc. MyoD1 gene expression is under epigenetic control and is regulated by HDAC and acetylation activity in the nucleus. MyoD1 is a 45kDa protein (320aa) with distinct function motifs. MyoD1 is expressed in muscle, liver, cancer cells, testis.
The MyoD1-selective antibodies were generated against peptide taken form with in the C0-terminal end of the protein between the aa acid (280-320). The MyoD1-selective antibodies are affinity purified over immobilized antigen based affinity chromatography which yielded high affinity epitope-specific antibodies. The MyoD1 antibodies label a protein of approximately 42-45kDa on Western blot using Western blot positive control for MyoD1(PC-MyoD1). MyoD1-selective antibodies are also available in FITC conjugated preparations for confocal, IHC, IF and immunocytochemical analyses. FabGennix Inc. will also conjugate antibodies to many other fluorescent probes upon request at a nominal charge. Limited quantities of antigenic blocking peptide and western blot positive controls are also available (inquire before ordering). FabGennix Inc. has an inventory of more than 1100 well characterized antibodies for research and diagnostic applications, for a complete listing please visit www.FabGennix.com.
Immunogen: Synthetic peptide selected from human MyoD1protein taken from within the C-terminal region (between aa 280-320) of MyoD1. The peptide sequence is conserved in several other species. The selected peptide was post-synthetically modified to achieve highest antigenicity before coupling to carrier protein using hetero bifunctional cross linker for immunogen preparation.
Concentration: Affinity purified MyoD1-101AP antibody (0.57-0.60mg/ml) was stabilized in antibody stabilization buffer for long-term storage.
Applications: MyoD1-101AP antibody was tested in ELISA and western blotting applications at 1:500 dilution using PC-MyoD1 samples. Antibody dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions. Application of this antibody in other protocols has not yet tested. WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d. Investigators using this antibody in protocols other than listed above can request a complimentary sample of this antibody.
Reactivity: The antibody labels an apparent MW of 42 kDa band in PC-MyoD1 samples and in mouse muscle C2C12 cell.
Protocols: Standard protocol for various applications (WB, IMM, IHC) of this antibody can be obtained upon request. The specification sheet for OC-STAMP antibody will be supplied with each product. FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.
Storage: The antiserum is supplied in antibody stabilization buffer with 0.02% azide. The purified antibodies are isolated from immobilized antigen affinity column. The affinity pure antibodies are supplied as 0.5-1 mg/ml IgG in antibody stabilization buffer. For long-term storage of keep at -20oC. FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DiluOBuffer). Working solutions of antibodies in DiluOBuffer should be filtered through 0.45m filter after every use for long-term storage.
Notes: Briefly centrifuge to collect liquid, heat or boil PC-FALP tube for 1-2 minutes to dissolve any precipitate before use. This product is “ready-to-use” for electrophoresis. After thawing store at room temperature, Repeated freezing and thawing may result in appearance of higher MW immunoreactive bands.
New Reagents: Now you can recycle your western blots (nitrocellulose, supported membranes and PVFD membranes) by using our StripObuffer (Cat FGI-1989). Each stripping is guaranteed to give better signal (up to 8 stripping). No strong pungent smell of reducing agents or heating required. Block in 5X diluObuffer and you are ready for blotting with a new antibody
