Anti-PKC 抗体 [EPR16794]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PKC antibody [EPR16794] (AB179521)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC alpha+beta1+beta2 with ab179521 at 1/100 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 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 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab179521 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 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 1/500 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] (AB179521)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human breast cancer is observed. Counter stained with Hematoxylin.

Negative 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] (AB179521)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse liver is observed. Counter stained with Hematoxylin.

Negative 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] (AB179521)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on rat testis is observed. Counter stained with Hematoxylin.

Negative 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.

• IP

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Immunoprecipitation - Anti-PKC antibody [EPR16794] (AB179521)

PKC alpha+beta1+beta2 was immunoprecipitated from 1mg of Mouse brain whole cell lysate with ab179521 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab179521 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : Mouse brain whole cell lysate 10μg (Input).
Lane 2 : ab179521 IP in Mouse brain whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab179521 in Mouse brain whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-PKC antibody [EPR16794] (ab179521)

Predicted band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16794] (AB179521)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

ab179521 weakly recognizes PKC gamma, and is only evident at long exposure times.

All lanes:

Western blot - Anti-PKC antibody [EPR16794] (ab179521) at 1/2000 dilution

All lanes:

Active human PKC gamma full length protein at 0.02 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 77 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16794] (AB179521)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

Human fetal kidney lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 77 kDa

Observed band size: 78 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16794] (AB179521)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-PKC antibody [EPR16794] (ab179521) at 1/2000 dilution

Lane 1:

Active human PKC alpha full length protein at 0.02 µg

Lane 2:

Active human PKC beta 1 full length protein at 0.02 µg

Lane 3:

Active human PKC beta 2 full length protein at 0.02 µg

Lane 4:

Active human PKC gamma full length protein at 0.02 µg

Lane 5:

Active human PKC delta full length protein at 0.02 µg

Lane 6:

Active human PKC eta full length protein at 0.02 µg

Lane 7:

Active human PKC epsilon full length protein at 0.02 µg

Lane 8:

Active human PKC theta full length protein at 0.02 µg

Lane 9:

Active human PKC mu full length protein at 0.02 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 77 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16794] (AB179521)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution

Lane 1:

L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate at 10 µg

Lane 2:

Rat thymus lysate at 10 µg

Lane 3:

Mouse thymus lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 27 kDa,77 kDa

Observed band size: 27 kDa,78 kDa

false

• WB

Unknown

Western blot - Anti-PKC antibody [EPR16794] (AB179521)

Recombinant PKC isoforms were tested for reactivity by western blot, and are marked as positive (+) or negative (-).

All lanes:

Western blot - Anti-PKC antibody [EPR16794] (ab179521)

Predicted band size: 77 kDa

false