Anti-Myogenin antibody [EPR4789]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myogenin antibody [EPR4789] (AB124800)

Immunohistochemical analysis of formalin fixed paraffin embedded human rhabdomyosarcoma labelling myogenin with ab124800 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab124800 Anti-Myogenin antibody [EPR4789] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myogenin antibody [EPR4789] (AB124800)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human rhabdomyosarcoma tissue sections labeling Myogenin with Purified ab124800 at 1 : 500 dilution (1.08 µg/ml). Heat mediated antigen retrieval was performed using using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myogenin antibody [EPR4789] (AB124800)

Unpurified ab124800, at a 1/100 dilution, staining Myogenin in paraffin embedded Human rhabdomyosarcoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Myogenin antibody [EPR4789] (AB124800)

Blocking and diluting buffer : 5% NFDM/TBST.

This antibody has low affinity on skeletal muscle samples. Please increase protein or antibody amount to get satisfied results.
Myogenin expression is upregulated in C2C12 cells during differentiation process induced by horse serum treatment (PMID : 26563778).

All lanes:

Western blot - Anti-Myogenin antibody [EPR4789] (ab124800) at 0.5 µg/mL

Lane 1:

Human fetal skeletal muscle lysates at 15 µg

Lane 2:

Rat fetal skeletal muscle lysates at 15 µg

Lane 3:

Mouse fetal skeletal muscle lysates at 15 µg

Lane 4:

C2C12 (Mouse myoblasts myoblast) whole cell lysates at 15 µg

Lane 5:

C2C12 (Mouse myoblasts myoblast) treated with 2% horse serum whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 25 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Myogenin antibody [EPR4789] (AB124800)

Immunofluorescence staining of Myogenin using ab124800 in ab277612, which were differentiated for 3 days post induction.

The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab124800 at 0.5 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.

The antibody ab124800 gave comparable results using MeOH fixation (100%, 5 min).