All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/2000 dilution

Lane 1 : Active human PKC alpha full length protein
Lane 2 : Active human PKC beta 1 full length protein
Lane 3 : Active human PKC beta 2 full length protein
Lane 4 : Active human PKC gamma full length protein
Lane 5 : Active human PKC delta full length protein
Lane 6 : Active human PKC eta full length protein
Lane 7 : Active human PKC epsilon full length protein
Lane 8 : Active human PKC theta full length protein
Lane 9 : 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 kDa
Observed band size: 103 kDa why is the actual band size different from the predicted?


Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

Active human PKC alpha full length protein (ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein (ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein (ab60841) contains aa1-673 with GST-tag; Active human PKC gamma full length protein (ab60842) contains aa1-677 with GST-tag; Active human PKC delta full length protein (ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein (ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein (ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein (ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein (ab60873) contains aa1-912 with GST-tag.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and weak cytoplasm staining on epithelial cells of Human colon 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.

 

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasm staining on K562 cell line. The nuclear counter stain 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: ab195039 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 K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PKC beta 1 with ab195039 at 1/200 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.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasm staining on HeLa cell line. The nuclear counter stain 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: ab195039 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.

 

All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/10000 dilution

Lane 1 : Human fetal brain whole cell lysate
Lane 2 : L-929 (Mouse connective tissue fibroblast cells) whole cell lysate
Lane 3 : C2C12 (Mouse myoblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 77 kDa
Observed band size: 40,77 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/10000 dilution

Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 77 kDa
Observed band size: 40,77 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 77 kDa
Observed band size: 40,77 kDa why is the actual band size different from the predicted?


Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

All lanes : Anti-PKC beta 1 antibody [EPR18512] (ab195039) at 1/2000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 77 kDa
Observed band size: 40,77 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The peptide immunogen corresponds to the C terminal region of human PKC beta 1. The antibody recognizes the full length (~77KD) and the catalytic domain (~40KD) of PKC beta 1 (Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331.

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining on cancer cells of colon cancer 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.

 

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm 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 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.

 

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling PKC beta 1 with ab195039 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus and cytoplasm staining on lymphocytes of rat spleen 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.