Anti-MATH2/NEUROD6 antibody [EPR28660-23]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse NeuroD6 expression vector containing a Myc-His tag. (B) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a Myc-His-tagged mouse NeuroD6 construct (image A). No staining on HEK-293T transfected with empty plasmid (image B). The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cardiac muscle (PMID : 7545978). The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat lung. The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse lung (PMID : 7545978). The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat cardiac muscle. The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum (PMID : 7545978). The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling MATH2/NEUROD6 with ab315812 at 1/4000 (0.133 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum. The section was incubated with ab315812 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

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Immunocytochemistry/ Immunofluorescence - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling MATH2/NEUROD6 with ab315812 at 1/50 (10.66 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 nuclear staining in mouse primary neurons.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4ug/ml) dilution (Red). 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labelling MATH2/NEUROD6 with ab315812 at 1/50 (10.66 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 nuclear staining in rat primary neuron.
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4ug/ml) dilution (Red). 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling MATH2/NEUROD6 with ab315812 at 1/50 dilution (1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

MATH2/NEUROD6 was immunoprecipitated from 0.35 mg Mouse P5 cerebral cortex tissue lysate with ab315812 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315812 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (ab315812) at 1/1000 dilution

Lane 1:

Mouse P5 cerebral cortex tissue lysate

Lane 2:

ab315812 at 1/30 IP in Mouse P5 cerebral cortex tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315812 in Mouse P5 cerebral cortex tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

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

Negative control : heart, lung (PMID : 7545978, 7744035)

The identity of the lower MW band at approximately 25 kDa is unknown.

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

All lanes:

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (ab315812) at 1/1000 dilution

Lane 1:

Mouse P5 cerebral cortex tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse lung tissue lysate at 20 µg

Lane 4:

Rat P5 hippocampus tissue lysate at 20 µg

Lane 5:

Rat heart tissue lysate at 20 µg

Lane 6:

Rat lung tissue lysate at 20 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 1/100000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

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

Negative control : kidney (PMID : 7545978, 7744035)

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

All lanes:

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (ab315812) at 1/1000 dilution

Lane 1:

Rat cerebral cortex tissue lysate at 20 µg

Lane 2:

Rat P5 brain tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 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 1/100000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

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

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

Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (ab315812) at 1/1000 dilution

Lane 1:

Mouse brain nuclear fraction at 20 µg

Lane 2:

Mouse brain non-nuclear fraction at 20 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 1/100000 dilution

Observed band size: 48 kDa,36 kDa,15 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (AB315812)

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

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-MATH2/NEUROD6 antibody [EPR28660-23] (ab315812) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse NEUROD6 expression vector containing a myc-His-tag®, whole cell lysate at 20 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 1/100000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 37s