Invitrogen

Myosin 4 Monoclonal Antibody (MF20), Alexa Fluor™ 488, eBioscience™

Product Image Gallery

Myosin 4 Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry of differentiated C2C12 cells using 5 µg/mL of Mouse IgG2b Isotype Control Alexa Fluor® 488 (Product # 53-4732-80) (left) or 5 µg/mL Anti-Myosin Heavy Chain Alexa Fluor® 488 (right). Myosin is positive in a subpopulation of differentiated cells. Nuclei are counterstained with DAPI.

Myosin 4 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure 1 TAK-242 reduces LPS-induced atrophy of C2C12 myotubes. ( A,B ) Western blot analysis ( A ) and quantification ( B ) of MyHC expression in C2C12 myotubes treated for 48 h with vehicle, LPS (1 mug/mL), or LPS and TAK-242 (1 muM). Data in ( B ) were normalised to beta-tubulin protein levels, and the ratio in vehicle-treated control cells was set to 1. N = 5/group. Full-length blots are presented in Supplementary Figure S4. ( C ) Representative immunofluorescence staining of MyHC in C2C12 myotubes treated as described for ( A,B ). Scale bar = 100 mum. ( D-F ) Distribution of myotube widths ( D ), mean myotube width ( E ), and fusion index ( F ) of C2C12 cells treated as described in ( A,B ). Myotube width was measured as described in the Methods. N = 97-114. The fusion index was calculated from five randomly selected fields as described in the Methods. For all panels, data are presented as the mean +- s.e.m. ***p < 0.001, **p < 0.01, *p < 0.05 vs vehicle control; ### p < 0.001, ## p < 0.01 vs LPS-treated group. P-values were derived from one-way ANOVA followed by Tukey's honest significant difference test or Kruskal-Wallis test followed by Dunn's post hoc tests with Bonferroni correction.

Myosin 4 Antibody in Immunohistochemistry (IHC) — Fig. 8 Identification and quantitation of the different cell types undergoing PCD during heart development. a Sagittal section of a transgenic heart at E12.5 depicting PCD cells. PCD cells were defined by condensed nuclei, a clear ring-like membrane sA5-YFP accumulation, and/or cCasp3 + signal. b Quantitation of PCD cells (white arrow in a). c Sagittal section of a transgenic heart at E12.5 depicting bodies of PCD cells. PCD Bodies were defined as smaller particles (2-4 um) without nuclear signal and positive for either sA5-YFP or cCasp3 or both. d Quantitation of PCD bodies (white arrow in c ). e - j Co-stainings in sections of transgenic embryonic hearts (E13.5) stained for cardiomyocyte (CMs, alpha-actinin, and MF20), endothelial cell (ECs, PECAM), leukocyte (LCs, CD45), smooth muscle cell (SMCs, ASMAC), and mesenchymal cell (MCs, vimentin) markers. k Quantitation of the different cell types that undergo PCD during heart development. Hoechst nuclear staining and sA5-YFP secreted human Annexin V-yellow fluorescent protein. Bars are 5 um ( a , c , e , f ), 10 um ( g , j ), and 20 um ( h , i ). Data are shown as mean +- s.e.m.,*** P < 0.001 ANOVA with Bonferroni post hoc test; n = 7-13

Myosin 4 Antibody in Western Blot (WB) — Fig. 6 In vitro administration of the pan-TAM tyrosine kinase inhibitor BMS-777607 impairs myogenesis of C2C12 myoblast cells. a mRNA expression levels of Tyro3, Axl, and Mer during the differentiation of C2C12 myoblast cells determined by qRT-PCR. b Protein expression levels of Axl and myosin heavy chain 4 (MYHC4) in differentiating C2C12 myoblasts in the presence or absence of 1 muM BMS-777607 determined by Western blot analysis. alpha-Tubulin was used as a loading control. One representative blot of three is shown. c Alterations in the number of viable C2C12 cells grown in growth medium in the presence or absence of 1 muM BMS-777607 determined by PrestoBlue staining. d Percent of cells in G2/M phase grown in growth medium in the presence or absence of 1 muM BMS-777607 as indicated by anti-phospho-histone H3 (Ser10) and DAPI co-staining (at least 20 HPF were analyzed). Arrows point to the anti-phospho-histone H3 positive nuclei. e Percent of PI-positive cells grown in growth medium in the presence or absence of 1 muM BMS-777607. f Alterations in the number of viable C2C12 cells grown in differentiation medium in the presence or absence of 1 muM BMS-777607 determined by PrestoBlue staining. BMS-777607 was added when the medium was changed to differentiation medium. g Representative light microscopic images of C2C12 myoblasts differentiated for 6 days in the absence or presence of 1 muM BMS-777607 after staining dead cells with propidium iodide and quantification of propidium

Myosin 4 Antibody in Flow Cytometry (Flow) — Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Flow Cytometry

Product Details

53-6503-82

Applications	Tested Dilution	Publications
Western Blot (WB)	-	View 4 publications 4 publications
Immunohistochemistry (IHC)	-	View 4 publications 4 publications
Immunohistochemistry (Paraffin) (IHC (P))	-	View 1 publication 1 publication
Immunohistochemistry (Frozen) (IHC (F))	Assay-Dependent	-
Immunocytochemistry (ICC/IF)	5 µg/mL	View 11 publications 11 publications
Flow Cytometry (Flow)	-	View 2 publications 2 publications

Product Specifications
Species Reactivity	Bovine, Dog, Chicken, Chimpanzee, Cat, Guinea pig, Human, Mouse, Non-human primate, Rabbit, Rat
Published species	Human, Mouse
Host/Isotype	Mouse / IgG2b, kappa
Recommended Isotype Control	Mouse IgG2b kappa Isotype Control (eBMG2b), Alexa Fluor™ 488, eBioscience™
Class	Monoclonal
Type	Antibody
Clone	MF20
Conjugate	Alexa Fluor™ 488 eFluor 660 Unconjugated
Excitation/Emission Profile	View spectra
Form	Liquid
Concentration	0.5 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2
Contains	0.09% sodium azide
Storage conditions	4° C, store in dark, DO NOT FREEZE!
RRID	AB_10671272

Product Specific Information

Description: This MF20 monoclonal antibody recognizes the heavy chain of myosin II, specificially the light meromyosin portion, in cardiac and skeletal muscle of vertebrates. Myosin II is composed of two heavy chains and four light chains. The 220-kDa myosin heavy chain exists as four different isoforms due to alternative splicing. Myosins interact with actin and hydrolyze ATP to function in muscle contraction, cytokinesis, and phagocytosis.

The MF20 has been shown to react to myosin from a variety of mammalian, avian and amphibian species, including rat, mouse, human, chicken, zebrafish, and dog.

Applications Reported: This MF20 antibody has been reported for use in immunohistochemical staining, immunocytochemistry, and immunohistochemical staining of frozen tissue sections.

Applications Tested: This MF20 antibody has been tested by immunocytochemistry on fixed and permeabilized C2C12 cells that were differentiated for 2 days in low serum medium prior to staining. Staining can be performed using less than or equal to 5 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.

Target Information

Target protein includes a fast myosin isoform expressed in skeletal muscle and has function involved in Muscle contraction.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

TAM kinase signaling is indispensable for proper skeletal muscle regeneration in mice.

Cell death & disease

Al-Zaeed N,Budai Z,Szondy Z,Sarang Z

53-6503-82 was used in Immunohistochemistry to identify Axl as a survival and growth receptor in the mouse myoblasts, and reveals the contribution of TAM kinase-mediated signaling to the skeletal muscle regeneration both in macrophages and in myoblasts.

Sat Jun 12 00:00:00 EDT 2021

TAM kinase signaling is indispensable for proper skeletal muscle regeneration in mice.

Cell death & disease

Al-Zaeed N,Budai Z,Szondy Z,Sarang Z

Sat Jun 12 00:00:00 EDT 2021

Data on the proliferation and differentiation of C2C12 myoblast treated with branched-chain ketoacid dehydrogenase kinase inhibitor.

Data in brief

Sato Y,Tate H,Yoshizawa F,Sato Y

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Western Blot

Sat Aug 01 00:00:00 EDT 2020

Comprehensive histological investigation of age-related changes in dermal extracellular matrix and muscle fibers in the upper lip vermilion.

International journal of cosmetic science

Gomi T,Imamura T

53-6503 was used in Immunohistochemistry-immunofluorescence to collect and study histological and molecular data on age-dependent alterations in the upper lip vermillion.

Sat Aug 01 00:00:00 EDT 2020

In vivo detection of programmed cell death during mouse heart development.

Cell death and differentiation

Martínez-Lagunas K,Yamaguchi Y,Becker C,Geisen C,DeRuiter MC,Miura M,Fleischmann BK,Hesse M

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Immunohistochemistry

Wed Apr 01 00:00:00 EDT 2020

TAK-242, a specific inhibitor of Toll-like receptor 4 signalling, prevents endotoxemia-induced skeletal muscle wasting in mice.

Scientific reports

Ono Y,Maejima Y,Saito M,Sakamoto K,Horita S,Shimomura K,Inoue S,Kotani J

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Western Blot

Mon Jan 20 00:00:00 EST 2020

TAK-242, a specific inhibitor of Toll-like receptor 4 signalling, prevents endotoxemia-induced skeletal muscle wasting in mice.

Scientific reports

Ono Y,Maejima Y,Saito M,Sakamoto K,Horita S,Shimomura K,Inoue S,Kotani J

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Western Blot

Mon Jan 20 00:00:00 EST 2020

Ternary Aligned Nanofibers of RGD Peptide-Displaying M13 Bacteriophage/PLGA/Graphene Oxide for Facilitated Myogenesis.

Nanotheranostics

Shin YC,Kim C,Song SJ,Jun S,Kim CS,Hong SW,Hyon SH,Han DW,Oh JW

53-6503 was used in Immunocytochemistry-immunoflourescence to fabricate poly(lactic-co-glycolic acid) nanofibre sheets decorated with graphene oxide and RGD peptid, as biofunctional scaffolds.

Wed Nov 20 00:00:00 EST 2019

Assessment of different strategies for scalable production and proliferation of human myoblasts.

Cell proliferation

Chua MJ,Yildirim ED,Tan JE,Chua YB,Low SC,Ding SLS,Li CW,Jiang Z,Teh BT,Yu K,Shyh-Chang N

53-6503 was used in Immunocytochemistry-immunoflourescence to show that, while all the strategies above are suboptimal at generating bona fide human myoblasts that can both proliferate and differentiate robustly, chemically defined hESC-monolayer-myoblasts show the most promise in differentiation potential.

Wed May 01 00:00:00 EDT 2019

Peptidyl-prolyl cis-trans isomerase NIMA interacting 1 regulates skeletal muscle fusion through structural modification of Smad3 in the linker region.

Journal of cellular physiology

Islam R,Yoon H,Shin HR,Bae HS,Kim BS,Yoon WJ,Woo KM,Baek JH,Lee YS,Ryoo HM

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Immunocytochemistry

Sat Dec 01 00:00:00 EST 2018

Classification of C2C12 cells at differentiation by convolutional neural network of deep learning using phase contrast images.

Human cell

Niioka H,Asatani S,Yoshimura A,Ohigashi H,Tagawa S,Miyake J

53-6503 was used in Immunocytochemistry to develop the convolutional neural network algorithm for automatic cellular differentiation recognition of the myogenic C12C12 cell line.

Mon Jan 01 00:00:00 EST 2018

An Engineered Optogenetic Switch for Spatiotemporal Control of Gene Expression, Cell Differentiation, and Tissue Morphogenesis.

ACS synthetic biology

Polstein LR,Juhas M,Hanna G,Bursac N,Gersbach CA

53-6503 was used in Immunocytochemistry-immunofluorescence to apply optogenetics to control cell differentiation and new tissue formation.

Fri Nov 17 00:00:00 EST 2017

NOTCH1-Dependent Nitric Oxide Signaling Deficiency in Hypoplastic Left Heart Syndrome Revealed Through Patient-Specific Phenotypes Detected in Bioengineered Cardiogenesis.

Stem cells (Dayton, Ohio)

Hrstka SC,Li X,Nelson TJ

53-6503 was used in Immunocytochemistry-immunoflourescence to study a mechanistic basis through which NOTCH1 contributes to heart development.

Sat Apr 01 00:00:00 EDT 2017

Natural underlying mtDNA heteroplasmy as a potential source of intra-person hiPSC variability.

The EMBO journal

Perales-Clemente E,Cook AN,Evans JM,Roellinger S,Secreto F,Emmanuele V,Oglesbee D,Mootha VK,Hirano M,Schon EA,Terzic A,Nelson TJ

53-6503 was used in Flow cytometry/Cell sorting to characterise the mitochondrial DNA of human induced pluripotent stem cells for use as a selection criteria.

Thu Sep 15 00:00:00 EDT 2016

Targeted Ablation of Periostin-Expressing Activated Fibroblasts Prevents Adverse Cardiac Remodeling in Mice.

Circulation research

Kaur H,Takefuji M,Ngai CY,Carvalho J,Bayer J,Wietelmann A,Poetsch A,Hoelper S,Conway SJ,Möllmann H,Looso M,Troidl C,Offermanns S,Wettschureck N

53-6503 was used in Immunohistochemistry to study the effect of activated cardiac fibroblast ablation on cardiac remodeling.

Fri Jun 10 00:00:00 EDT 2016

Ultrastructure of medial rectus muscles in patients with intermittent exotropia.

Eye (London, England)

Yao J,Wang X,Ren H,Liu G,Lu P

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Western Blot

Fri Jan 01 00:00:00 EST 2016

Single-molecule analysis of myocyte differentiation reveals bimodal lineage commitment.

Integrative biology : quantitative biosciences from nano to macro

Gibson TM,Gersbach CA

53-6503 was used in Immunofluorescence to characterise the differentiation of single skeletal muscle cells.

Mon Jun 01 00:00:00 EDT 2015

Single-molecule analysis of myocyte differentiation reveals bimodal lineage commitment.

Integrative biology : quantitative biosciences from nano to macro

Gibson TM,Gersbach CA

53-6503 was used in Immunofluorescence to characterise the differentiation of single skeletal muscle cells.

Mon Jun 01 00:00:00 EDT 2015

Inhibition of TGFβ signaling increases direct conversion of fibroblasts to induced cardiomyocytes.

PloS one

Ifkovits JL,Addis RC,Epstein JA,Gearhart JD

Published figure using Myosin 4 monoclonal antibody (Product # 53-6503-82) in Immunocytochemistry

Tue Jan 20 00:00:00 EST 2015

iPS cell-derived cardiogenicity is hindered by sustained integration of reprogramming transgenes.

Circulation. Cardiovascular genetics

Martinez-Fernandez A,Nelson TJ,Reyes S,Alekseev AE,Secreto F,Perez-Terzic C,Beraldi R,Sung HK,Nagy A,Terzic A

Wed Oct 01 00:00:00 EDT 2014

iPS cell-derived cardiogenicity is hindered by sustained integration of reprogramming transgenes.

Circulation. Cardiovascular genetics

Martinez-Fernandez A,Nelson TJ,Reyes S,Alekseev AE,Secreto F,Perez-Terzic C,Beraldi R,Sung HK,Nagy A,Terzic A

Wed Oct 01 00:00:00 EDT 2014

iPS cell-derived cardiogenicity is hindered by sustained integration of reprogramming transgenes.

Circulation. Cardiovascular genetics

Martinez-Fernandez A,Nelson TJ,Reyes S,Alekseev AE,Secreto F,Perez-Terzic C,Beraldi R,Sung HK,Nagy A,Terzic A

Wed Oct 01 00:00:00 EDT 2014

Optimization of direct fibroblast reprogramming to cardiomyocytes using calcium activity as a functional measure of success.

Journal of molecular and cellular cardiology

Addis RC,Ifkovits JL,Pinto F,Kellam LD,Esteso P,Rentschler S,Christoforou N,Epstein JA,Gearhart JD

53-6503 was used in Immunohistochemistry to conclude that HNGMT produces iCMs more efficiently than previously published methods.

Mon Jul 01 00:00:00 EDT 2013

Optimization of direct fibroblast reprogramming to cardiomyocytes using calcium activity as a functional measure of success.

Journal of molecular and cellular cardiology

Addis RC,Ifkovits JL,Pinto F,Kellam LD,Esteso P,Rentschler S,Christoforou N,Epstein JA,Gearhart JD

53-6503 was used in Immunohistochemistry to conclude that HNGMT produces iCMs more efficiently than previously published methods.

Mon Jul 01 00:00:00 EDT 2013

Bioinformatics

Protein Aliases: 2B myosin heavy chain; fast myosin heavy chain 2B; myh4; MyHC IIb isoform; MyHC-2b; MyHC-IIb; Myosin heavy chain 2b; myosin heavy chain 2X; Myosin heavy chain 4; Myosin heavy chain IIb; myosin heavy chain type IIb; Myosin heavy chain, skeletal muscle; Myosin heavy chain, skeletal muscle, fetal; Myosin heavy chain, skeletal muscle, juvenile; myosin, heavy chain 4, skeletal muscle; myosin, heavy polypeptide 4, skeletal muscle; Myosin-4; unnamed protein product

Gene Aliases: AI506973; MHC; MHC2B; Minimsc; Minmus; MM; MYH-2B; MYH1; MYH2B; MYH4; MyHC-2B; MyHC-IIb; Myhsf

UniProt ID: (Human) Q9Y623, (Dog) Q076A5, (Rabbit) Q28641, (Mouse) Q5SX39, (Rat) Q29RW1

Entrez Gene ID: (Human) 4622, (Bovine) 408020, (Dog) 479502, (Rabbit) 100009548, (Rabbit) 100125991, (Guinea pig) 100726456, (Mouse) 17884, (Rat) 360543