Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 4 hr 30 min
- Sample type: Adherent cells, Purified protein, Suspension cells, Tissue Extracts
Product namePKC Kinase Activity Assay Kit
Intra-assay Sample n Mean SD CV% Sample < 10% Inter-assay Sample n Mean SD CV% Sample < 10%
Sample typeAdherent cells, Suspension cells, Tissue Extracts, Purified protein
Assay typeEnzyme activity
Assay time4h 30m
PKC Kinase Activity Assay Kit (ab139437) is a non-radioactive assay providing a safe, rapid and reliable method for the screening of inhibitors or activators of PKC and for quantitating the activity of PKC in purified or partially purified enzyme preparations. This kit is based on a solid phase enzyme-linked immuno-absorbent assay (ELISA) that utilizes a specific synthetic peptide as a substrate for PKC and a polyclonal antibody that recognizes the phosphorylated form of the substrate. The assay is designed for the analysis of PKC activity in the solution phase. For the measurement of PKC in partially purified, purified, or crude enzyme preparations from any species.
The kit offers the following advantages:
1. Safe - non-radioactive measurement of kinase activity.
2. Flexible - kinetic and end-point options available.
3. Fast - results in < 4.5 hours.
4. Efficient - only 30 μl diluted sample needed per well.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 20X Wash Buffer 1 x 30ml Active PKC 1 x 30µl Antibody Dilution Buffer 1 x 10ml Anti-Rabbit IgG: HRP Conjugate 1 x 20µl ATP 1 x 2mg Kinase Assay Dilution Buffer 1 x 10ml PKC Phosphospecific Substrate Antibody 1 x 5ml PKC Substrate Microtiter Plate 1 unit Stop Solution 2 1 x 10ml TMB Substrate 1 x 10ml
RelevanceProtein Kinase C (PKC) isoforms are serine/threonine kinases involved in signal transduction pathways that govern a wide range of physiological processes including differentiation, proliferation, gene expression, brain function, membrane transport and the organization of cytoskeletal and extracellular matrix proteins. Increasing evidence from studies using in vitro and in vivo systems points to PKC as a key regulator of critical cell cycle transitions, including cell cycle entry and exit and the G1 and G2 checkpoints. PKC-mediated control of these transitions can be negative or positive, depending on the timing of PKC activation during the cell cycle and on the specific PKC isozymes involved. There have been at least 12 different PKC isoforms identified in humans to date, including alpha, beta I, beta II, gamma, delta, epsilon, zeta, eta, theta, iota, lambda, and mu.
- Phorbol 12-myristate 13-acetate (PMA), PKC activator (ab120297)
- GF 109203X hydrochloride, ATP-competitive PKC inhibitor (ab120844)
- Go 6976, PKC inhibitor (ab141413)
- 1-O-Hexadecyl-2-O-acetyl-sn-glycerol, Protein Kinase C Inhibitor (ab141550)
- HBDDE, PKCalpha and PKCgamma inhibitor (ab141573)
- Calphostin C, PKC inhibitor (ab142407)
- Phorbol-12,13-dibutyrate, PKC activator (ab142636)
1.5 x e7 THP-1 cells were incubated with 100 nM GF109203X (GF; ab144264) or Ro31-8220 mesylate (Ro; ab120374) for 30 minutes prior to activation with 10 μg x mL-1 PMA (Sigma) for 4 hours. Control cells were left without inhibitors or PMA. Cells were lysed in 1 mL of lysis buffer, and 30 μL were tested for PKC activity (duplicates; +/- SD).
Titration of ab139437 (duplicates; +/- SD).
Signal from active PKC with background signal subtracted (duplicates; +/- SD).
ab139437 has been referenced in 23 publications.
- Julku UH et al. Prolyl Oligopeptidase Regulates Dopamine Transporter Oligomerization and Phosphorylation in a PKC- and ERK-Independent Manner. Int J Mol Sci 22:N/A (2021). PubMed: 33579026
- Grubišic V et al. Enteric Glia Modulate Macrophage Phenotype and Visceral Sensitivity following Inflammation. Cell Rep 32:108100 (2020). PubMed: 32905782
- Farrow MA et al. Small Molecule Inhibitor Screen Reveals Calcium Channel Signaling as a Mechanistic Mediator of Clostridium difficile TcdB-Induced Necrosis. ACS Chem Biol 15:1212-1221 (2020). PubMed: 31909964
- Tapia R et al. Enteropathogenic Escherichia coli (EPEC) Recruitment of PAR Polarity Protein Atypical PKC? to Pedestals and Cell-Cell Contacts Precedes Disruption of Tight Junctions in Intestinal Epithelial Cells. Int J Mol Sci 21:N/A (2020). PubMed: 31947656
- Domínguez-García S et al. A novel PKC activating molecule promotes neuroblast differentiation and delivery of newborn neurons in brain injuries. Cell Death Dis 11:262 (2020). PubMed: 32321920