Invitrogen

MYOD Monoclonal Antibody (5.8A)

Advanced Verification

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

4 Published Figures

View all (23) MYOD antibodies

Create Citation

Questions & Answers

Create Citation

Questions & Answers

Product Image Gallery

MYOD Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of MYOD was performed using 70% confluent log phase differentiated C2C12 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MYOD monoclonal antibody (Product # MA1-41017) at 5ug/mL dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 conjugate (Product # A28175) at a dilution of 1:2000 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: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image. Panel e represents undifferentiated C2C12 cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

MYOD Antibody in Immunohistochemistry (IHC) — Immunohistochemical analysis of human tissues using a the MYOD monoclonal antibody (Product # MA1-41017). A. Rhabdomyosaroma (nuclei are stained), B. Lymphoma (staining is absent)

MYOD Antibody in Western Blot (WB) — Western blot was performed using Anti-MYOD Monoclonal Antibody (Product # MA1-41017) and a 40kDa band corresponding to MYOD was observed across cell lines tested and was observed to increase upon C2C12 differentiated to 3 and 7 days respectively. Modified whole cell extracts (1% SDS) (30 µg lysate) of C2C12 (Lane 1), C2C12 differentiated to myotubes (3 days) (Lane 2) and C2C12 differentiated to myotubes (7 days) (Lane 3) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 (1ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

MYOD Antibody in Western Blot (WB) — Western blot analysis for MyoD expression in various small round cell tumor lines using 1 µg/mL of a MYOD monoclonal antibody (Product # MA1-41017). The antibody only reacts with a band of approx. 45 kD in the rhabdomyosarcoma cell line (Rh30, lane 1) but was negative against the primitive neuroectodermal (PFSK-1A, lane 2), lymphoma (EB2, lane 3), neuroblastoma (SK-N-SH, lane 4), and Ewing's sarcoma (SJSA-1, lane 5) cell lines.

MYOD Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cells lines tested owing to their inherent genetic constitution. Relative expression of MYOD was observed in C2C12 cells differentiated to myotubes (3 and 7 days) in comparison to undifferentiated C2C12 using Anti-MYOD Monoclonal Antibody (Product # MA1-41017) in Western Blot. {RE}

MYOD Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Immunofluorescence analysis using MYOD Monoclonal Antibody (Product # MA5-27909) showed expression of MYOD in C2C12 differentiated to myotubes as compared to undifferentiated C2C12 cells. {RE}

MYOD Antibody — Fig. 13 Ch25 overexpression in TA muscle inhibits the expression of myogenic markers and induces apoptosis. (A-C) Cryosections of TA muscles injected with lentivirus overexpressing 25-hydroxylase gene or lenti virus carrying empty vector were immunostained with MyoD, myogenin or Pax7, and laminin antibodies, and nuclei were stained with DAPI. (D) TUNEL staining in cryosections of TA muscles injected with lentivirus that overexpress 25-hydroxylase gene or lenti virus carrying empty vector. Fig. 13

MYOD Antibody — Fig. 5 Ang II infusion leads to increased apoptosis and inhibition of myogenic differentiation in C57BL/6 but not in TNFR1KO mice. (A) TUNEL staining in cryosections of gastrocnemius muscles from ang II infused or pair-fed sham infused C57BL/6 or TNFR1KO mice. (B-D) MyoD, myogenin and Pax 7 staining in cryosections of gastrocnemius muscles from ang II infused or pair-fed sham infused C57BL/6 or TNFR1KO mice. The sections were immunostained with laminin and MyoD, myogenin or Pax7 antibodies, and nuclei were stained with DAPI. The pictures show MyoD, myogenin and Pax7 positive cells (green, indicated by white arrows) and basement membrane outlined by laminin staining (red). Fig. 5

MYOD Antibody — Figure 1 Rapamycin blocks C2C12 differentiation. (a) The order of appearance of myotube differentiation markers over the course of 15 days in differentiation media only. This is representative of three independent experiments. (b) Differences in differentation marker transcripts when treated with DMSO (vehicle) or 500 nM rapamycin for 9 days, throughout the differentiation protocol. The most recent rapamycin administration was 1 day prior to cell lysis. These data represent the average of three wells, from a representative experiment (n = 3). Transcripts from both (a) and (b) were measured by qRT-PCR and normalized to Gapdh . (c) Representative western blot analysis of C2C12 cells treated for 4 h with Differentiation Media (2% Horse Serum; HS) or left in growth media (10% FBS) in the presence of 500 nM Rapamycin or DMSO. (d) Quantification of the blots in c (n = 4 independent experiments). Protein phosphorylation is presented as the intensity of the phosphospecific antibody band relative to that proteins total protein band. (e) Images of morphological changes in C2C12 myoblasts in response to 10 days of DMSO or rapamycin treatment througout the differentiation protocol (500 nM). Asterisks indicate p < 0.05. Data represents mean +/- standard error of the mean.

MYOD Antibody — Published figure using MYOD monoclonal antibody (Product # MA1-41017)

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

5 µg/mL

View 4 publications 4 publications

Immunofluorescence (IF)

5 µg/mL

View 2 publications 2 publications

Immunohistochemistry (Frozen) (IHC (F))

1:10-1:500

View 1 publication 1 publication

Immunoprecipitation (IP)

1 µg/mL

-

Western Blot (WB)

1 µg/mL

View 4 publications 4 publications

Miscellaneous PubMed (Misc)

-

View 1 publication 1 publication

Immunohistochemistry (IHC)

-

View 2 publications 2 publications

Product Specifications

Species

Human, Mouse, Rat

Published species

Artificial Control, Human, Mouse, Rat

Expression System

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

5.8A

Immunogen

Recombinant mouse MyoD.

Conjugate

Unconjugated

Form

Liquid

Concentration

1.0 mg/mL

Purification

Protein G

Storage buffer

PBS

Contains

0.02% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2282434

Product Specific Information

The epitope of this antibody was mapped to a region within AA 180-189 of mouse MyoD.

Suggested positive control: Rh-30.

Target Information

The Myogenic determination gene (MyoD) was first identified by the virtue of its ability to convert embryonic mouse fibroblast cells to muscle cells. It was subsequently shown that forced expression of MyoD (human homolog is myf 3) gene in a wide variety of normal and neoplastic cells could either convert the cells to muscle cells or activate a set of the otherwise transcriptionally inactive muscle-specific genes in these cells. The regulatory domain of the MyoD gene product lies within a 70 amino acid region and comprises a basic DNA binding motif and a helix-loop-helix (HLH) dimerization motif. Subsequent studies identified three other genes whose products shared sequence homology for the basic HLH domain of MyoD. These are; myf5, myogenin (human homolog is myf4) and myf6 (also known as MRF4 and herculin).

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

Bioinformatics

Protein Aliases: bHLHc1; Class C basic helix-loop-helix protein 1; Myf-3; Myoblast determination protein 1; Myogenic factor 3

Gene Aliases: AI503393; BHLHC1; MYF3; MYOD; Myod-1; MYOD1; PUM

UniProt ID: (Human) P15172, (Mouse) P10085, (Rat) Q02346

Entrez Gene ID: (Human) 4654, (Mouse) 17927, (Rat) 337868

Molecular Function: basic helix-loop-helix transcription factor nucleic acid binding transcription factor

regulation of alternative mRNA splicing, via spliceosome regulation of transcription from RNA polymerase II promoter transcription from RNA polymerase II promoter protein phosphorylation muscle organ development myoblast fate determination skeletal muscle tissue development myoblast fusion cellular response to starvation myotube cell development myotube differentiation involved in skeletal muscle regeneration skeletal muscle cell differentiation muscle cell fate commitment positive regulation of skeletal muscle tissue regeneration regulation of RNA splicing skeletal muscle fiber adaptation histone H3 acetylation histone H4 acetylation positive regulation of myoblast differentiation positive regulation of transcription from RNA polymerase II promoter skeletal muscle fiber development positive regulation of skeletal muscle fiber development positive regulation of muscle cell differentiation cellular response to glucocorticoid stimulus cellular response to estradiol stimulus cellular response to oxygen levels positive regulation of myoblast fusion negative regulation of myoblast proliferation transcription, DNA-templated regulation of transcription, DNA-templated multicellular organism development myotube differentiation cell differentiation skeletal muscle tissue regeneration myoblast differentiation positive regulation of transcription, DNA-templated striated muscle cell differentiation cardiac muscle cell differentiation skeletal muscle atrophy response to muscle stretch

Process(es)

RNA polymerase II regulatory region sequence-specific DNA binding RNA polymerase II core promoter proximal region sequence-specific DNA binding core promoter binding transcriptional activator activity, RNA polymerase II transcription regulatory region sequence-specific binding chromatin binding transcription factor activity, RNA polymerase II distal enhancer sequence-specific binding transcription coactivator activity protein binding transcription factor binding chromatin DNA binding ubiquitin protein ligase binding nuclear hormone receptor binding protein heterodimerization activity E-box binding DNA binding transcription factor activity, sequence-specific DNA binding enzyme binding sequence-specific DNA binding protein dimerization activity