PKC Kinase Activity Assay Kit (ab139437)
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
Overview
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Product name
PKC Kinase Activity Assay Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Sample Inter-assay Sample n Mean SD CV% Sample -
Sample type
Adherent cells, Suspension cells, Tissue Extracts, Purified protein -
Assay type
Enzyme activity -
Assay time
4h 30m -
Product overview
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. Efficient - only 30 μl diluted sample needed per well.
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Platform
Microplate reader
Properties
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Storage instructions
Please 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 -
Research areas
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Relevance
Protein 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.
Images
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Functional assay: PKC Kinase Activity Assay Kit (ab139437)
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).
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schematic diagram of the assay This image is courtesy of Fitzpatrick, Daniel P. (2020) "A Mechanistic Study on the Non-genotoxic Carcinogenicity of the Food Contaminant Semicarbazide," Sure-J: Science Undergraduate Research Journal: Vol. 2: Iss. 1, Article 2 -
Functional assay: PKC Kinase Activity Assay Kit (ab139437)Titration of ab139437 (duplicates; +/- SD).
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Functional assay: PKC Kinase Activity Assay Kit (ab139437)Signal from active PKC with background signal subtracted (duplicates; +/- SD).