Invitrogen

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

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

15 Published Figures

26 References

View all (8) Myosin 4 antibodies

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Datasheet

Protocols

Questions & Answers

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Myosin 4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of MYH4 was performed using 70% confluent log phase C2C12 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Myosin 4 Monoclonal Antibody (MF20), Alexa Fluor 488, eBioscience™ (Product # 53-6503-82) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 647 (Product # A32787, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Blue) was stained withAlexa Fluor™ 488 Phalloidin (Product # A12379, 1:300). Panel d represents the merged image showing Cytoskeleton localization. Panel e represents C2c12 undifferentiated. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Myosin 4 Antibody in Western Blot (WB) — Western blot was performed using Myosin 4 Monoclonal Antibody (MF20), Alexa Fluor 488, eBioscience™ (Product # 53-6503-82) and a ~230 kDa band corresponding to MYH4 was observed across all tissues. Whole cell extracts (30 µg lysate) of Mouse Heart (Lane 1), Rat Heart (Lane 2), Mouse Skeletal Muscle (Lane 3), Rat Skeletal Muscle (Lane 4), Mouse Brain (Lane 5), Rat Brain (Lane 6), Mouse Liver (Lane 7) were electrophoresed using NuPAGE™ 3 to 8%, Tris-Acetate, 1.0–1.5 mm, Mini Protein Gels (Product # EA03752BOX), 12 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

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

Myosin 4 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissue models owing to their inherent genetic constitution. Relative expression of MYH4 was observed between Mouse Heart, Rat Heart, Mouse Skeletal Muscle, Rat Skeletal Muscle which are positive models and Mouse Brain, Rat Brain and Mouse Liver which are negetive models using Myosin 4 Monoclonal Antibody (MF20), Alexa Fluor 488, eBioscience™ (Product # 53-6503-82) in Western Blot.

Product Details

53-6503-82

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000

View 5 publications 5 publications

Immunohistochemistry (IHC)

View 5 publications 5 publications

Immunohistochemistry (Paraffin) (IHC (P))

View 1 publication 1 publication

Immunohistochemistry (Frozen) (IHC (F))

Assay-Dependent

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1:100

View 12 publications 12 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

Excitation/Emission Max

499/520 nm 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

Impaired Skeletal Muscle Development and Regeneration in Transglutaminase 2 Knockout Mice.

Cells

Budai Z,Al-Zaeed N,Szentesi P,Halász H,Csernoch L,Szondy Z,Sarang Z

53-6503-82 was used in Western Blot, Immunohistochemistry (Frozen) to test the involvement of transglutaminase 2 (TG2) in muscle development and regeneration using a full-body TG2 knockout mouse strain.

Tue Nov 09 00:00:00 EST 2021

Impaired Skeletal Muscle Development and Regeneration in Transglutaminase 2 Knockout Mice.

Cells

Budai Z,Al-Zaeed N,Szentesi P,Halász H,Csernoch L,Szondy Z,Sarang Z

Tue Nov 09 00:00:00 EST 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

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

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

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

Sat Jun 12 00:00:00 EDT 2021

hiPSC-Derived Neurons Provide a Robust and Physiologically Relevant In Vitro Platform to Test Botulinum Neurotoxins.

Frontiers in pharmacology

Lamotte JD,Roqueviere S,Gautier H,Raban E,Bouré C,Fonfria E,Krupp J,Nicoleau C

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

Tue Feb 02 00:00:00 EST 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 Immunofluorescence

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 Immunofluorescence

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

Micropatterned substrates with physiological stiffness promote cell maturation and Pompe disease phenotype in human induced pluripotent stem cell-derived skeletal myocytes.

Biotechnology and bioengineering

Jiwlawat N,Lynch EM,Napiwocki BN,Stempien A,Ashton RS,Kamp TJ,Crone WC,Suzuki M

53-6503-82 was used in Immunohistochemistry to demonstrate that human induced pluripotent stem cell (iPSC)-derived myogenic progenitors formed highly-aligned myotubes and contracted when seeded on two-dimensional micropatterned platforms.

Sun Sep 01 00:00:00 EDT 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

Function(s)

muscle contraction response to activity actin filament-based movement muscle filament sliding ATP metabolic process metabolic process response to gravity response to muscle activity response to muscle stretch muscle structure development

Process(es)

microfilament motor activity double-stranded RNA binding actin binding calmodulin binding ATP binding ATPase activity motor activity actin filament binding nucleotide binding molecular_function

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

Product Details

53-6503-82

Western Blot (WB)

Immunohistochemistry (IHC)

Immunohistochemistry (Paraffin) (IHC (P))

Immunohistochemistry (Frozen) (IHC (F))

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Species Reactivity

Published species

Host/Isotype

Recommended Isotype Control

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

Performance Guarantee

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