Adherent cell cultures were fixed with 4% PFA, permeabilized with 0.2% Triton-X in PBS, and incubated with primary antibodies: myogenin (M-225) from Santa Cruz (Dallas, TX, USA), α-MyHC (Developmental Studies Hybridoma Bank), and α-Laminin (Sigma). Alexa-488 and Alexa-555 conjugated secondary antibodies that matched the primary antibodies were used at 1:1,000 in PBS (Invitrogen). Nuclei were counterstained with DAPI (Sigma-Aldrich). Images were obtained using an Axioplan2 microscope (Carl Zeiss, Oberkochen, Germany), a ×20 NA 0.75 plan Apochromat (ω/0.17; Carl Zeiss) objective, and a digital Axiocam camera (Carl Zeiss). Digital images were captured using Axiovision (Carl Zeiss) and were processed with Photoshop (Adobe, San Jose, CA, USA).
Myogenin m 225
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Myogenin (M-225) is a mouse monoclonal antibody targeted against the myogenin protein, a member of the MyoD family of transcription factors. It is used in research applications for the detection and analysis of myogenin expression.
Automatically generated - may contain errors
Lab products found in correlation
α laminin, by Merck Group (1 mentions)
Axioplan 2 microscope, by Zeiss (1 mentions)
Axiocam camera, by Zeiss (1 mentions)
Axiovision, by Zeiss (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Phosphorylated erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Total erk1 2, by Cell Signaling Technology (1 mentions)
Phosphorylated akt serine 473, by Cell Signaling Technology (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Imaging densitometer, by Bio-Rad (1 mentions)
P38 map kinase, by Cell Signaling Technology (1 mentions)
Myod m318, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene fluoride (pvdf), by Bio-Rad (1 mentions)
3 protocols using myogenin m 225
1
Immunofluorescence Imaging of Adherent Cells
2
Immunoblotting of Muscle Proteins
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Immunoblotting was performed as previously described [9 (link)], and the antibodies included MHC (MF20; DSHB; 1:1000), myogenin (M-225; Santa Cruz; #sc-576; 1:1000), phosphorylated Akt (serine473; Cell Signaling; #9271; 1:1000), total Akt (Cell Signaling; #9272; 1:1000), phosphorylated ERK1/2 (Thr202/Tyr204; Cell Signaling; #9101; 1:1000); total ERK1/2 (Cell Signaling; #9102; 1:1000) and β-actin (Sigma Aldrich; #A2228; 1:1000).
3
Western Blot Analysis of Muscle Proteins
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For the analysis of protein expression, muscle samples were separated on a 15% polyacrylamide gel and the proteins transferred to PVDF (Biorad, Ca, USA). The membrane was probed with primary antibodies diluted in Tris buffered saline (TBS) with 0.1% (v/v) Tween-20 and 0.5% (w/v) milk powder. Membranes were incubated with horseradish peroxidase-conjugated secondary antibodies before detection of antibody binding by chemiluminescence (Amersham). The membranes were reprobed for anti-b-actin to check protein loading. Band intensity was measured using a BioRad imaging densitometer and quantified using Molecular Analyst software.
Antibodies used were as follows; mouse monoclonal antibodies against Pax7 at 1:1000 myogenin F5D at 1:500 (DSHB) and MyoD 5.8A at 1:1000 (BD Biosciences). Rabbit polyclonal antibodies against Myf5 at 1:1000, MyoD M-318 at 1:1000 and myogenin M-225 (Santa Cruz, TX, USA) at 1:500. p38MAPKinase at 1:1000 and phosphorylated p38MAPKinase (Cell Signalling, USA) at 1:1000. Goat polyclonal antibody against Pax3 (Abcam, MA, USA) at 1:2000. Secondary antibodies were either; anti-mouse IgG (Sigma) at 1:2000, anti-rabbit IgG (Sigma) at 1:2000 or anti-goat IgG (Santa Cruz) at 1:2000.
Antibodies used were as follows; mouse monoclonal antibodies against Pax7 at 1:1000 myogenin F5D at 1:500 (DSHB) and MyoD 5.8A at 1:1000 (BD Biosciences). Rabbit polyclonal antibodies against Myf5 at 1:1000, MyoD M-318 at 1:1000 and myogenin M-225 (Santa Cruz, TX, USA) at 1:500. p38MAPKinase at 1:1000 and phosphorylated p38MAPKinase (Cell Signalling, USA) at 1:1000. Goat polyclonal antibody against Pax3 (Abcam, MA, USA) at 1:2000. Secondary antibodies were either; anti-mouse IgG (Sigma) at 1:2000, anti-rabbit IgG (Sigma) at 1:2000 or anti-goat IgG (Santa Cruz) at 1:2000.
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