重组Anti-PKC gamma抗体[EPR28643-68] - BSA and Azide free (ab317316)

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse cerebellum tissue staining PKC gamma with ab317315 at a 1:5000 (0.1 μg/ml) dilution, KCNA2 with ab316324 at 1:4000 (0.124 μg/ml) dilution and KCND2 with ab307710 at a 1:2000 (0.266 μg/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A: merged staining of anti-PKC gamma (green; Opal™520), anti-KCNA2 (magenta; Opal™690) and anti-KCND2 (gray; Opal™570) on mouse cerebellum.
Panel B: anti-PKC gamma staining the Purkinje cells and neuropil in mouse cerebellum.
Panel C: anti-KCNA2 staining the basket cells in mouse cerebellum.
Panel D: anti-KCND2 staining the granule cell layer in mouse cerebellum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining: in the order of ab317315, ab313624 and ab307710 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat cerebellum tissue staining PKC gamma with ab317315 at a 1:5000 (0.1 μg/ml) dilution, KCNA2 with ab316324 at 1:4000 (0.124 μg/ml) dilution and KCND2 with ab307710 at a 1:2000 (0.266 μg/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A: merged staining of anti-PKC gamma (green; Opal™520), anti-KCNA2 (magenta; Opal™690) and anti-KCND2 (gray; Opal™570) on rat cerebellum.
Panel B: anti-PKC gamma staining the Purkinje cells and neuropil in rat cerebellum.
Panel C: anti-KCNA2 staining the basket cells in rat cerebellum.
Panel D: anti-KCND2 staining the granule cell layer in rat cerebellum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining: in the order of ab317315, ab313624 and ab307710 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

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

Negative control: spleen(PMID:1556149)

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

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

Negative control: spleen, liver, heart(PMID:1556149)

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling PKC gamma with ab317315 at 1/100 (5.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human cerebellum.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling PKC gamma with ab317315 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse cerebellum.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling PKC gamma with ab317315 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in rat cerebellum.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling PKC gamma with ab317315 at 1/100 (5.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: No staining in human liver.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling PKC gamma with ab317315 at 1/100 (5.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: No staining in human spleen.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling PKC gamma with ab317315 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: No staining in mouse liver.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PKC gamma with ab317315 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: No staining in mouse spleen.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling PKC gamma with ab317315 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: No staining in rat liver.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling PKC gamma with ab317315 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: No staining in rat spleen.
The section was incubated with ab317315 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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh frozen) tissue labeling PKC gamma with ab317315 at 1/500 (1.01 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Panel A: merged staining of anti-PKC gamma(ab317315, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on mouse cerebellum.
Panel B: anti-PKC gamma stained on mouse cerebellum.
Panel C: anti-NeuN stained in neurons of mouse cerebellum.
Panel D: anti-GFAP stained in astrocytes of mouse cerebellum.
The section was incubated in two rounds of staining: in the order of ab317315 and ab190565, ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse heart (fresh frozen) tissue labeling PKC gamma with ab317315 at 1/500 (1.01 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Negative control: confocal image showing no staining on mouse heart. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317315 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (fresh frozen) tissue labeling PKC gamma with ab317315 at 1/500 (1.01 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Panel A: merged staining of anti-PKC gamma(ab317315, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on rat cerebellum.
Panel B: anti-PKC gamma stained on rat cerebellum.
Panel C: anti-NeuN stained in neurons of rat cerebellum.
Panel D: anti-GFAP stained in astrocytes of rat cerebellum.
The section was incubated in two rounds of staining: in the order of ab317315 and ab190565, ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat heart (fresh frozen) tissue labeling PKC gamma with ab317315 at 1/500 (1.01 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Negative control: confocal image showing no staining on rat heart (PMID: 1556149). The nuclear counterstain was DAPI (Blue). The section was incubated with ab317315 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse hippocampal neuron cells labelling PKC gamma with ab317315 at 1/200 (2.525 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 hippocampal neuron (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).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4 (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.

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

PKC gamma was immunoprecipitated from 0.35 mg Human cerebellum tissue lysate with ab317315 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317315 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Human cerebellum tissue lysate
Lane 2: ab317315 IP in Human cerebellum tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab317315 in human cerebellum tissue lysate

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

Exposure time: 180 seconds.

This data was developed using ab317315, the same antibody clone in a different buffer formulation.

Dot blot analysis of PKC gamma using ab317315 at 1:1000 (0.505 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1:100,000 dilution.

Lane 1: Human PKC gamma peptide

Lane 2: His-tagged Human PKC alpha protein

Exposure time: 180 seconds.

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

This antibody doesn't cross react with Human PKC alpha.