Anti-Myosin antibody [A4.1025] (ab37484) at 0.1614 µg/ml + Human skeletal muscle tissue lysate at 20 µg

Secondary
Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 223 kDa
Observed band size: 223 kDa


Exposure time: 3 seconds

The extra bands under 223KDa were mainly caused by degradation (PMID: 3295257).

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling Myosin (MYH1/2) with ab37484 at 0.807 µg/ml, followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on human skeletal muscle. The section was incubated with ab37484 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse skeletal muscle tissue labeling Myosin (MYH1/2) with ab37484 at 16.14 µg/ml followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse skeletal muscle is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells labelling Myosin (MYH1/2) with ab37484 at 3.228 µg/ml, followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (2µg/ml) (Green). Confocal image showing cytoplasmic staining in differentiated (treated) C2C12 cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (2.5µg/ml), followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/500 dilution (4µg/ml) (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (2µg/ml).

All lanes : Anti-Myosin antibody [A4.1025] (ab37484) at 0.807 µg/ml

Lane 1 : Mouse skeletal muscle tissue lysate
Lane 2 : Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 223 kDa
Observed band size: 223 kDa

Exposure times: Lane 1: 3 secs; Lane 2: 1 sec.

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-Myosin antibody [A4.1025] (ab37484) at 0.807 µg/ml

Lane 1 : C2C12 (mouse myoblasts) whole cell lysate
Lane 2 : Differentiated C2C12 whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 223 kDa
Observed band size: 223 kDa


Exposure time: 3 seconds

The expression profile observed is consistent with what has been described in the literature (PMID: 26010876).

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling Myosin (MYH1/2) with ab37484 at 0.807 µg/ml, followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on human cardiac muscle. The section was incubated with ab37484 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue labeling Myosin (MYH1/2) with ab37484 at 0.807 µg/ml, followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on mouse skeletal muscle. The section was incubated with ab37484 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue labeling Myosin (MYH1/2) with ab37484 at 0.807 µg/ml, followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on rat  skeletal muscle. The section was incubated with ab37484 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat skeletal muscle tissue labeling Myosin (MYH1/2) with ab37484 at 16.14 µg/ml followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat skeletal muscle is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).