Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RD (human muscle multinucleated spindle-shaped cell) cells labelling Sarcomeric Alpha Actinin with ab317695 at 1/500 (1.05 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic and membranous staining in RD cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Sarcomeric Alpha Actinin with ab317695 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse skeletal muscle. The section was incubated with ab317695 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary cardiomyocyte cells labelling Sarcomeric Alpha Actinin with ab317695 at 1/500 (1.05 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing positive staining in mouse primary cardiomyocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab9465 Anti-Sarcomeric Alpha Actinin mouse monoclonal antibody was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab317695 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab9465 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling Sarcomeric Alpha Actinin with ab317695 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cardiac muscle. The section was incubated with ab317695 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling Sarcomeric Alpha Actinin with ab317695 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat skeletal muscle. The section was incubated with ab317695 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblast) cells labelling Sarcomeric Alpha Actinin with ab317695 at 1/500 (1.05 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic and membranous staining in differentiated C2C12 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Sarcomeric Alpha Actinin with ab317695 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver. The section was incubated with ab317695 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Sarcomeric Alpha Actinin with ab317695 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat liver. The section was incubated with ab317695 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary cardiomyocyte cells labelling Sarcomeric Alpha Actinin with ab317695 at 1/500 (1.05 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing positive staining in rat primary cardiomyocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab9465 Anti-Sarcomeric Alpha Actinin mouse monoclonal antibody was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab317695 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab9465 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Negative control : liver, pancreas, placenta (PMID : 11440986).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (ab317695) at 1/1000 dilution

Lane 1:

Human heart tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 50 µg

Lane 3:

Human liver tissue lysate at 50 µg

Lane 4:

Human pancreas tissue lysate at 50 µg

Lane 5:

Human placenta tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 103 kDa,36 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Dot blot analysis of Sarcomeric Alpha Actinin using ab317695 at 1 : 1000 (0.525 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lane 1 : His-tagged human ACTN2 recombinant protein
Lane 2 : His-tagged human ACTN4 recombinant protein
Lane 3 : His-tagged human ACTN1 recombinant protein
Lane 4 : His-tagged human ACTN3 recombinant protein

Anti-His tag antibody (1 : 10000) as total protein control.

This antibody does not cross-react with human ACTN1, human ACTN3 and human ACTN4.

All lanes:

Dot Blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (ab317695) at 1/1000 dilution

Lane 1:

His-tagged human ACTN2 recombinant protein

Lane 2:

His-tagged human ACTN4 recombinant protein

Lane 3:

His-tagged human ACTN1 recombinant protein

Lane 4:

His-tagged human ACTN3 recombinant protein

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Negative control : liver (PMID : 11440986).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (ab317695) at 1/1000 dilution

Lane 1:

Rat skeletal muscle tissue lysate at 50 µg

Lane 2:

Rat heart tissue lysate at 50 µg

Lane 3:

Rat liver tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 103 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (AB317695)

Negative control : liver, placenta (PMID : 11440986).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 180 seconds, Lanes 3-6 : 26 seconds

All lanes:

Western blot - Anti-Sarcomeric Alpha Actinin antibody [EPR28776-83] (ab317695) at 1/1000 dilution

Lane 1:

Untreated C2C12 (mouse myoblast) whole cell lysate at 50 µg

Lane 2:

C2C12 differentiation to muscle for 5 days whole cell lysate at 50 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 50 µg

Lane 4:

Mouse heart tissue lysate at 50 µg

Lane 5:

Mouse liver tissue lysate at 50 µg

Lane 6:

Mouse placenta tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 103 kDa,36 kDa

false