Analysis of Myosin Heavy Chain in human embryonic stem cell (hESC)-derived cardiomyocytes. hESC- derived cardiomyocytes (Evans laboratory, UC San Diego) were disassociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885), and stained with PE Mouse Anti-Myosin Heavy Chain (MHC) and Alexa Fluor® 647 Mouse Anti-Cardiac Troponin I (cTnI) (Cat. No. 554409). The cells were first gated on light scatter properties and then analyzed for MHC and cTnI expression. Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
Analysis of Myosin Heavy Chain in differentiated C2C12 mouse myoblast cell line. C2C12 cells (ATCC: CRL-1772) that were differentiated for 3 days and then disassociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash buffer I (Cat. No. 557885), and stained with PE Mouse Anti-Myosin Heavy Chain (MHC). The cells were first gated on light scatter properties and then analyzed for MHC expression. Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
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Analysis of Myosin Heavy Chain in human embryonic stem cell (hESC)-derived cardiomyocytes. hESC- derived cardiomyocytes (Evans laboratory, UC San Diego) were disassociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885), and stained with PE Mouse Anti-Myosin Heavy Chain (MHC) and Alexa Fluor® 647 Mouse Anti-Cardiac Troponin I (cTnI) (Cat. No. 554409). The cells were first gated on light scatter properties and then analyzed for MHC and cTnI expression. Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
Analysis of Myosin Heavy Chain in differentiated C2C12 mouse myoblast cell line. C2C12 cells (ATCC: CRL-1772) that were differentiated for 3 days and then disassociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash buffer I (Cat. No. 557885), and stained with PE Mouse Anti-Myosin Heavy Chain (MHC). The cells were first gated on light scatter properties and then analyzed for MHC expression. Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
Product Details
BD Pharmingen™
Mouse (QC Testing), Human (Tested in Development), Rat, Dog, Chicken, Frog, Zebrafish (Reported)
Mouse C3H, also known as C3H/He, C3H/Bi IgG2b, κ
Chicken pectoralis myosin
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2738795
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
Alexa Fluor® 647 Mouse Anti-Cardiac Troponin I RUO
Size
50 Tests
Cat No.
564409
564408 Rev. 1
Antibody Details
MF20
The MF20 monoclonal antibody specifically binds to the heavy chain component of skeletal muscle myosin or myosin II. Skeletal muscle myosin is composed of 2 heavy chains and 4 light chains. Myosin Heavy chain is a 220-kDa protein that interacts with actin in the ATP-dependent mechanism of muscle contraction. This antibody is reported to cross react with human, mouse, rat, chicken, dog, frog, and zebrafish.
564408 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
564408 Rev.1
Citations & References
Development References (5)
-
Bader D, Masaki T, Fischman DA. Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro. J Cell Biol. 1982; 95(3):763-770. (Immunogen). View Reference
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Dambrot C, Passier R, Atsma D, Mummery CL. Cardiomyocyte differentiation of pluripotent stem cells and their use as cardiac disease models. Biochem J. 2011; 434(1):25-35. (Biology). View Reference
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Lian X, Hsiao C, Wilson G, et al. Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A. 2012; 109(27):E1848-E1857. (Biology). View Reference
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López JE, Myagmar BE, Swigart PM, et al. β-myosin heavy chain is induced by pressure overload in a minor subpopulation of smaller mouse cardiac myocytes. Circ Res. 2011; 109(6):629-638. (Clone-specific). View Reference
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Travaglione S, Messina G, Fabbri A. Cytotoxic necrotizing factor 1 hinders skeletal muscle differentiation in vitro by perturbing the activation/deactivation balance of Rho GTPases. Cell Death Differ. 2005; 12(1):78-86. (Clone-specific). View Reference
564408 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
