Invitrogen

Myogenin Polyclonal Antibody

Advanced Verification

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

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Questions & Answers

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Myogenin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Myogenin was performed using 70% confluent log phase RD 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 Myogenin Rabbit Polyclonal Antibody (Product # PA5-78067) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) 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 showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Myogenin Antibody in Western Blot (WB) — Western blot was performed using Anti-Myogenin Polyclonal Antibody (Product # PA5-78067) and a 30 kDa band corresponding to MYOG was observed in RD except HEK-293 and U-2 OS which is reported to be negative. Modified whole cell extracts (1% SDS) (30 µg lysate) of RD (Lane 1), HEK-293 (Lane 2) and U-2 OS (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Myogenin Antibody in Western Blot (WB) — Western blot analysis of Myogenin in whole cell lysate using 30 µg of protein. Samples were separated with 12% SDS-PAGE and incubated with Myogenin polyclonal antibody (Product # PA5-78067) using a dilution of 1:1000.

Myogenin Antibody in ChIP assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous Myogenin protein using Myogenin Antibody: ChIP was performed using Anti-Myogenin Rabbit Polyclonal Antibody (Product # PA5-78067, 2.5 µg) on sheared chromatin from RD cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter region of DESMIN, NES and gene body of SIX1. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Myogenin Antibody in ChIP assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous Myogenin protein using Myogenin Antibody: ChIP was performed using Anti-Myogenin Rabbit Polyclonal Antibody (Product # PA5-78067, 2.5 µg) on sheared chromatin from RD cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter region of DESMIN, NES and gene body of SIX1. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Myogenin Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines tested owing to their inherent genetic constitution. Relative expression of MYOG was observed in RD in comparison to HEK-293 and U-2 OS using Anti-Myogenin Polyclonal Antibody (Product # PA5-78067) in Western Blot. {RE}

Product Details

PA5-78067

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	-
Immunocytochemistry (ICC/IF)	5 µg/mL	-
ChIP assay (ChIP)	2.5 µg/10^6 cells	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein encompassing a sequence within the center region of human Myogenin. The exact sequence is proprietary.
Conjugate	Unconjugated
Form	Liquid
Concentration	0.59 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol
Contains	0.025% Proclin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2736181

Product Specific Information

Positive Control: RD

Predicted Reactivity: Mouse (95%), Rat (95%), Pig (95%), Rabbit (96%), Sheep (92%), Bovine (92%)

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

Myogenin is a muscle-specific transcription factor that can induce myogenesis in a variety of cell types in tissue culture. It is a member of a large family of proteins related by sequence homology, the helix-loop-helix proteins. It is essential for the development of functional skeletal muscle.

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

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Bioinformatics

Protein Aliases: bHLHc3; Class C basic helix-loop-helix protein 3; Myf-4; Myogenic factor 4; Myogenic factor-4; Myogenin

Gene Aliases: BHLHC3; myf-4; MYF4; MYOG

UniProt ID: (Human) P15173

Entrez Gene ID: (Human) 4656

Molecular Function: basic helix-loop-helix transcription factor

ossification cell cycle skeletal muscle tissue development negative regulation of cell proliferation response to gravity positive regulation of myotube differentiation skeletal muscle atrophy positive regulation of muscle atrophy regulation of skeletal muscle satellite cell proliferation response to muscle activity involved in regulation of muscle adaptation response to electrical stimulus involved in regulation of muscle adaptation striated muscle atrophy response to denervation involved in regulation of muscle adaptation skeletal muscle cell differentiation muscle cell fate commitment mRNA transcription from RNA polymerase II promoter skeletal muscle tissue regeneration myoblast differentiation positive regulation of myoblast differentiation negative regulation of glycolytic process 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 positive regulation of cell cycle arrest cellular response to magnetism cellular response to lithium ion cellular response to retinoic acid cellular response to growth factor stimulus cellular response to estradiol stimulus regulation of myoblast fusion positive regulation of myoblast fusion positive regulation of oxidative phosphorylation

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 core promoter proximal region sequence-specific binding transcriptional activator activity, RNA polymerase II transcription regulatory region sequence-specific binding transcription factor activity, sequence-specific DNA binding transcription factor activity, RNA polymerase II distal enhancer sequence-specific binding protein binding chromatin DNA binding sequence-specific DNA binding protein heterodimerization activity E-box binding