Anti-PKC gamma antibody [EPR28643-68] (ab317315)

Rabbit Recombinant Monoclonal KPCG antibody. Suitable for mIHC, Dot, WB, IP, IHC-P, IHC-Fr, ICC/IF and reacts with Mouse, Rat, Human, Synthetic peptide - Human samples.

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Images

Immunohistochemistry (Frozen sections) - Anti-PKC gamma antibody [EPR28643-68] (AB317315), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	mIHC	Dot	WB	IP	IHC-P	IHC-Fr	ICC/IF	Flow Cyt (Intra)
Human	Expected	Expected	Tested	Tested	Tested	Expected	Expected	Not recommended
Mouse	Tested	Expected	Tested	Expected	Tested	Tested	Tested	Not recommended
Rat	Tested	Expected	Tested	Expected	Tested	Tested	Not recommended	Not recommended
Synthetic peptide - Human	Not recommended	Tested	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Rat	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/200	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat, Synthetic peptide - Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat, Synthetic peptide - Human	Dilution info -	Notes -

Target data

See full target information PRKCG

Function

Calcium-activated, phospholipid- and diacylglycerol (DAG)-dependent serine/threonine-protein kinase that plays diverse roles in neuronal cells and eye tissues, such as regulation of the neuronal receptors GRIA4/GLUR4 and GRIN1/NMDAR1, modulation of receptors and neuronal functions related to sensitivity to opiates, pain and alcohol, mediation of synaptic function and cell survival after ischemia, and inhibition of gap junction activity after oxidative stress. Binds and phosphorylates GRIA4/GLUR4 glutamate receptor and regulates its function by increasing plasma membrane-associated GRIA4 expression. In primary cerebellar neurons treated with the agonist 3,5-dihyidroxyphenylglycine, functions downstream of the metabotropic glutamate receptor GRM5/MGLUR5 and phosphorylates GRIN1/NMDAR1 receptor which plays a key role in synaptic plasticity, synaptogenesis, excitotoxicity, memory acquisition and learning. May be involved in the regulation of hippocampal long-term potentiation (LTP), but may be not necessary for the process of synaptic plasticity. May be involved in desensitization of mu-type opioid receptor-mediated G-protein activation in the spinal cord, and may be critical for the development and/or maintenance of morphine-induced reinforcing effects in the limbic forebrain. May modulate the functionality of mu-type-opioid receptors by participating in a signaling pathway which leads to the phosphorylation and degradation of opioid receptors. May also contributes to chronic morphine-induced changes in nociceptive processing. Plays a role in neuropathic pain mechanisms and contributes to the maintenance of the allodynia pain produced by peripheral inflammation. Plays an important role in initial sensitivity and tolerance to ethanol, by mediating the behavioral effects of ethanol as well as the effects of this drug on the GABA(A) receptors. During and after cerebral ischemia modulate neurotransmission and cell survival in synaptic membranes, and is involved in insulin-induced inhibition of necrosis, an important mechanism for minimizing ischemic injury. Required for the elimination of multiple climbing fibers during innervation of Purkinje cells in developing cerebellum. Is activated in lens epithelial cells upon hydrogen peroxide treatment, and phosphorylates connexin-43 (GJA1/CX43), resulting in disassembly of GJA1 gap junction plaques and inhibition of gap junction activity which could provide a protective effect against oxidative stress (By similarity). Phosphorylates p53/TP53 and promotes p53/TP53-dependent apoptosis in response to DNA damage. Involved in the phase resetting of the cerebral cortex circadian clock during temporally restricted feeding. Stabilizes the core clock component BMAL1 by interfering with its ubiquitination, thus suppressing its degradation, resulting in phase resetting of the cerebral cortex clock (By similarity). Phosphorylates and activates LRRK1, which phosphorylates RAB proteins involved in intracellular trafficking (PubMed:36040231).

Alternative names

PKCG, PRKCG, Protein kinase C gamma type, PKC-gamma

Recommended products

Rabbit Recombinant Monoclonal KPCG antibody. Suitable for mIHC, Dot, WB, IP, IHC-P, IHC-Fr, ICC/IF and reacts with Mouse, Rat, Human, Synthetic peptide - Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR28643-68
Purification technique: Affinity purification Protein A
Specificity: Unsuitable for rat ICC.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Protein kinase C gamma (PKC gamma) also known as PKCγ is a member of the protein kinase C (PKC) family. This protein has a molecular weight of approximately 78 kDa. Expressed mainly in the central nervous system PKC gamma participates in various signaling pathways. It exists as a serine/threonine kinase and plays a fundamental role in modulating cellular functions in neurons.

Biological function summary

PKC gamma acts in the regulation of neuronal signaling processes and is a component of the synaptic transmission machinery. It associates with other proteins to form complex structures that influence physiological processes. PKC gamma contributes significantly to the functions of the central nervous system by modulating synaptic plasticity which is essential for learning and memory.

Pathways

PKC gamma participates in the MAPK and PI3K signaling pathways which are critical in cell growth and survival. The protein interacts with other PKC isozymes and signaling molecules within these pathways playing a role in phosphorylating specific target proteins. These interactions facilitate the regulation of cell cycle progression and apoptosis indicating its importance in cellular homeostasis.

Associated diseases and disorders

Abnormal PKC gamma activity associates with spinocerebellar ataxia type 14 and addiction-related behavior. Spinocerebellar ataxia results from mutations affecting PKC gamma which disrupts its normal function and leads to neurodegeneration. Additionally PKC gamma’s interaction with other proteins such as GABAA receptors in addiction highlights potential therapeutic targets for treatment.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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20 product images

Immunohistochemistry (Frozen sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.
Multiplex immunohistochemistry - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Anti-SCN11A antibody [EPR28603-66] ab316324 at 1:4000 (0.124 µg/ml) dilution and KCND2 with Anti-Kv4.2/KCND2 antibody [EPR26384-89] 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, Anti-KCNA antibody [EPR26383-85] ab313624 and Anti-Kv4.2/KCND2 antibody [EPR26384-89] 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
Multiplex immunohistochemistry - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Anti-SCN11A antibody [EPR28603-66] ab316324 at 1:4000 (0.124 µg/ml) dilution and KCND2 with Anti-Kv4.2/KCND2 antibody [EPR26384-89] 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, Anti-KCNA antibody [EPR26383-85] ab313624 and Anti-Kv4.2/KCND2 antibody [EPR26384-89] 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
Western blot - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
Negative control: spleen, liver, heart(PMID:1556149)
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-PKC gamma antibody [EPR28643-68] (ab317315) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg with NFDM/TBST
Lane 2: Mouse cerebellum tissue lysate at 20 µg with NFDM/TBST
Lane 3: Mouse spleen tissue lysate at 20 µg with NFDM/TBST
Lane 4: Mouse liver tissue lysate at 20 µg with NFDM/TBST
Lane 5: Mouse heart tissue lysate at 20 µg with NFDM/TBST
Lane 6: Rat brain tissue lysate at 20 µg with NFDM/TBST
Lane 7: Rat cerebellum tissue lysate at 20 µg with NFDM/TBST
Lane 8: Rat spleen tissue lysate at 20 µg with NFDM/TBST
Lane 9: Rat liver tissue lysate at 20 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 80 kDa, 36 kDa
Exposure time: 10s
Western blot - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
Negative control: spleen(PMID:1556149)
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-PKC gamma antibody [EPR28643-68] (ab317315) at 1/1000 dilution
Lane 1: Human cerebellum tissue lysate at 20 µg with NFDM/TBST
Lane 2: Human hippocampus tissue lysate at 20 µg with NFDM/TBST
Lane 3: Human spleen tissue lysate at 20 µg with NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 80 kDa, 36 kDa
Exposure time: 37s
Dot Blot - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.
This antibody doesn't cross react with Human PKC alpha.
All lanes: Dot Blot - Anti-PKC gamma antibody [EPR28643-68] (ab317315) at 1/1000 dilution
Lane 1: Human PKC gamma peptide with NFDM/TBST
Lane 2: His-tagged Human PKC alpha protein with NFDM/TBST
Secondary
All lanes: Dot Blot at 1/100000 dilution
Exposure time: 180s
Immunohistochemistry (Frozen sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.
Immunohistochemistry (Frozen sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 (Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, grey) and anti-GFAP (Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] 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 Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.
Immunohistochemistry (Frozen sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 (Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, grey) and anti-GFAP (Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] 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 Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.
Immunocytochemistry/ Immunofluorescence - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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).
Anti-MAP2 antibody [HM-2] 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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
Immunoprecipitation - Anti-PKC gamma antibody [EPR28643-68] (ab317315)
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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab317315 in human cerebellum tissue lysate
All lanes: Immunoprecipitation - Anti-PKC gamma antibody [EPR28643-68] (ab317315) at 1/30 dilution
All lanes: Human cerebellum tissue lysate with NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s