Antibodies, Proteins, Kits and Reagents for Life Science

264 681 ₸ Product size

100 µl 264 681 ₸ 10 µl 107 212 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16794] to PKC
Suitable for: WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-PKC antibody [EPR16794]
See all PKC primary antibodies
Description

Rabbit monoclonal [EPR16794] to PKC
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Mouse
WB	Mouse Rat Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Active human PKC alpha full length protein; Active human PKC beta 1 full length protein; Active human PKC beta 2 full length protein; Active human PKC gamma full length protein; Human fetal brain and fetal kidney lysates; Jurkat, SH-SY5Y and L-929 whole cell lysates; Rat and mouse thymus lysates. IHC-P: Human breast cancer, Mouse liver and Rat testis tissues. ICC/IF: HeLa cells. IP: Mouse brain whole cell lysate.
General notes
Recombinant PKC isoforms were tested for reactivity to ab179521 by western blot, and are marked as positive (+) or negative (-).
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16794
Isotype

IgG
Research areas

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

All lanes : Anti-PKC antibody [EPR16794] (ab179521) 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, 78 kDa

Exposure time: 5 seconds

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

Anti-PKC antibody [EPR16794] (ab179521) at 1/2000 dilution + Active human PKC gamma 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 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.
Western blot - Anti-PKC antibody [EPR16794] (ab179521)

Recombinant PKC isoforms were tested for reactivity by western blot, and are marked as positive (+) or negative (-).
Western blot - Anti-PKC antibody [EPR16794] (ab179521)

All lanes : Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

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

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

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-PKC antibody [EPR16794] (ab179521)

All lanes : Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution

Lane 1 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
Lane 2 : Rat thymus lysate
Lane 3 : Mouse thymus lysate

Lysates/proteins at 10 µg per lane.

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

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

Blocking/Dilution buffer: 5% NFDM/TBST.
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.
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.
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.
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.
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.
Anti-PKC antibody [EPR16794] (ab179521)

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