Analytical Tools for Protein Kinase C Isozymes

Protein Kinase C (PKC), and its various isozymes, are regulatory proteins that modify the function of other proteins via phosphorlyation.

The PKC group target serine or threonine for phosphorylation to impact the function of proteins involved in biological processes and pathways including cell death, cell growth and division, differentiation and memory.

There are 10 different isozymes in the PKC group which are classified into 3 subgroups – novel, conventional or atypical. Which group a specific PKC isozyme is classified into depends on the structure of the proteins regulatory region.

Analysis tools which allow researchers to differentiate between different PKC isozymes are needed to research specific PKC pathways. Abcam provide a range of antibodies specific to different isozymes (including for their phosphorylated state), and inhibitors for each PKC isozyme subgroup:

Conventional Novel Atypical
  • PKC alpha
  • PKC beta
  • PKC delta
  • PKC epsilon
  • PKC zeta

Conventional

PKC enzymes of the ‘conventional’ subgroup have four domains with homologous structures (C1, C2, C3 and C4). They are activated by the binding of a phospholipid, phorbol or esters, alongside calcium and DAG (diacylglycerol).

Analytical tools that can be used against the conventional PKC enzymes (alpha and beta) include inhibitors and antibodies:

PKC Alpha

Immunohistochemical analysis of mouse small intestine stained with anti-PKC alpha [Y124] (ab32376).

Figure 1. Immunohistochemical analysis of mouse small intestine stained with anti-PKC alpha [Y124] (ab32376).

  • Go 6976 (ab141413) - Selective inhibitor of PKC alpha and PKC beta
  • HBDDE (ab141573) - Selective inhibitor of PKC alpha and PKC gamma
  • Phosphorylated PKC alpha antibodies
  • Anti-PKC alpha primary antibody [Y124] (ab32376) (knockout validated)

PKC Beta

Immunocytochemical analysis of K562 (human chronic myelogenous leukemia cells from bone marrow) cells labeled with anti-PKC beta 1 with (ab195039) at 1/100 followed by goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1,000 (green)

Figure 2. Immunocytochemical analysis of K562 (human chronic myelogenous leukemia cells from bone marrow) cells labeled with anti-PKC beta 1 with (ab195039) at 1/100 followed by goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1,000 (green)

  • LY 333531 hydrochloride - Selective inhibitor of PKC beta
  • Phosphorylated PKC beta antibodies
  • Anti-PKC beta 1 primary antibody [EPR18512] (ab195039)

Novel

PKC enzymes in the ‘novel’ subgroup are also activated by DAG, however they do not require calcium. The structure of this subgroups C2 domain is different to the conventional structure, with no calcium sensor.

Analytical tools that can be used against the novel PKC enzymes (delta and epsilon) include inhibitors and antibodies:

PKC Delta

Immunocytochemical analysis of HeLa cells labeled with anti-PKC delta (ab182126) at 1/100 followed by goat anti-rabbit Alexa Fluor® 488 (IgG) (ab150077) at 1/400 (green).

Figure 3. Immunocytochemical analysis of HeLa cells labeled with anti-PKC delta (ab182126) at 1/100 followed by goat anti-rabbit Alexa Fluor® 488 (IgG) (ab150077) at 1/400 (green).

  • Rottlerin - Inhibitor of PKC delta
  • Phosphorylated PKC delta antibodies
  • Anti-PKC delta primary antibody [EPR17075] (ab182126) (knockout validated)

PKC Epsilon

Immunohistochemical analysis of human endometrium tissue labeled with anti-PKC epsilon (ab124806) at 1/50.

Figure 4. Immunohistochemical analysis of human endometrium tissue labeled with anti-PKC epsilon (ab124806) at 1/50.

  • V1-2 - Inhibitor of PKC epsilon
  • Phosphorylated PKC epsilon antibodies
  • Anti-PKC epsilon primary antibody [EPR1482(2)] (ab124806)

Atypical

PKC enzymes of the ‘atypical’ subgroup do not require secondary activation as they have no C2 domain, and their C1 domain is non-functional.

Analytical tools that can be used against the atypical PKC enzymes (delta and epsilon) include inhibitors and antibodies:

PKC Zeta

Immunocytochemical analysis of human HeLa cells labeled with anti-PKC zeta (ab108970) at 1/150 followed by goat anti-rabbit IgG (Alexa Fluor® 488) at 1/2,000.

Figure 5. Immunocytochemical analysis of human HeLa cells labeled with anti-PKC zeta (ab108970) at 1/150 followed by goat anti-rabbit IgG (Alexa Fluor® 488) at 1/2,000.

  • Phosphorylated zeta antibodies
  • Anti-PKC zeta primary antibody [EP1490(2)] (ab108970) (knockout validated)

Pan PKC

For some experiments, researchers may want to look at all the PKC enzymes in a sample, with no specificity towards specific isozymes or subgroups. For this research there are inhibitors, antibodies and kits which target pan PKC:

Immunohistochemical analysis of mouse testis tissue labeled with anti-PKC antibody [EPR17368] (ab181558) at 1/1,000, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

Figure 6. Immunohistochemical analysis of mouse testis tissue labeled with anti-PKC antibody [EPR17368] (ab181558) at 1/1,000, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

  • Sotrastaurin - Inhibitor of PKC
  • Anti-PKC antibody [EPR17368] (ab181558) – for different isoforms
  • PKC Kinase Activity Assay Kit (ab139437)

Other PKC Isozyme Antibodies

  • Anti-PKC beta 2 antibody [Y125] (ab32026)
  • Anti-PKC gamma antibody [EPR2750] (ab108961)
  • Anti- PKC eta antibody [EPR18513] (ab179524)
  • Anti-PKC theta antibody [EPR1487(2)] (ab110728)
  • Anti-PKC iota antibody (ab154681)

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.


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Last updated: Apr 23, 2019 at 9:52 AM

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