All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

Lane 1 : Active human PKC alpha full length protein
Lane 2 : Active human PKC gamma full length protein

Lysates/proteins at 0.02 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 77, 78 kDa


Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

Active human PKC alpha full length protein (Catalog#ab55672) contains aa1-672 with GST-tag; Active human PKC gamma full length protein (Catalog#ab60842) contains aa1-677 with GST-tag.

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus and weak cytoplasmic staining on Human pancreatic acinar cells is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling PKC gamma with ab184746 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic and nuclear staining on SH-SY5Y cell line. 

The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab184746 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC gamma with ab184746 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

Lane 1 : Active human PKC beta 2 full length protein
Lane 2 : Active human PKC beta 1 full length protein
Lane 3 : Active human PKC delta full length protein
Lane 4 : Active human PKC eta full length protein
Lane 5 : Active human PKC epsilon full length protein
Lane 6 : Active human PKC theta full length protein
Lane 7 : Active human PKC mu full length protein

Lysates/proteins at 0.02 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 77, 78 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Active human PKC beta 1 full length protein(Catalog#ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein(Catalog#ab60841) contains aa1-673 with GST-tag; Active human PKC delta full length protein(Catalog#ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein(Catalog#ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein(Catalog#ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein(Catalog#ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein(Catalog#ab60873) contains aa1-912 with GST-tag.

Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution + Active human PKC beta 1 full length protein at 0.02 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 77, 78 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Active human PKC beta 1 full length protein (Catalog #ab60840) contains aa1-671 with GST-tag.

Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution + Human fetal brain lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 77, 78 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?


Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 77, 78 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?


Exposure time: 2 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (ab184746) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 6 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 77, 78 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?


Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC gamma with ab184746 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic and nuclear staining on HeLa cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab184746 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

Immunohistochemical analysis of paraffin-embedded cervical cancer tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus and weak cytoplasmic staining on cancer cells of cervix cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on epithelial cells of mouse kidney is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus and cytoplasmic staining on epithelial cells of rat kidney is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling PKC gamma with ab184746 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Weak cytoplasmic staining on neuron of Human cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.