Overview

  • Product name

    PKA Kinase Activity Assay Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Sample < 10%
    Inter-assay
    Sample n Mean SD CV%
    Sample <=12%
  • Sample type

    Adherent cells, Suspension cells, Tissue Extracts, Purified protein
  • Assay type

    Enzyme activity
  • Assay time

    4h 30m
  • Product overview

    PKA Kinase Activity Kit (ab139435) is a non-radioactive assay providing a safe, rapid and reliable method for the screening of inhibitors or activators of PKA and for quantitating the activity of PKA 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 PKA and a polyclonal antibody that recognizes the phosphorylated form of the substrate. The assay is designed for the analysis of PKA activity in the solution phase. This kit gives you the ability to an end-point or kinetic assay read-out in a convenient 96-well plate based assay and offers easy sample handling protocols.

    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

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer 1 x 30ml
    Active PKA 20 ng/µl 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
    PKA Phosphospecific Substrate Antibody 1 x 5ml
    PKA Substrate Microtiter Plate 1 unit
    Stop Solution 2 1 x 10ml
    TMB Substrate 1 x 10ml
  • Research areas

  • Relevance

    In cell biology, Protein kinase A (PKA) is a family of enzymes whose activity is dependent on cellular levels of cyclic AMP (cAMP). PKA is also known as cAMP-dependent protein kinase. Protein kinase A has several functions in the cell, including regulation of glycogen, sugar, and lipid metabolism.

Images

  • 1e7 3T3-L1 cells were treated for 15 minutes with 1 μM dexamethasone (ab120743) and 1 μg/mL insulin (ab123768) with the addition of 50 μM forskolin (ab120058), 0.5 mM IBMX (ab120840) or 0.1 mM 8-CPT-cAMP (ab120424). Cell extracts were prepared according to the protocol. Lysates were diluted 50-fold, and tested in duplicates (+/-SD).

  • Titration of positive control (active PKA), background signal subtracted (duplicates; +/- SD).

  • Titration of cAMP Protein Kinase (ab56268), background signal subtracted (duplicates; +/- SD).

Protocols

References

This product has been referenced in:

  • Shahrestanaki MK  et al. IPP-1 controls Akt/CREB phosphorylation extension in A2a adenosine receptor signaling cascade in MIN6 pancreatic ß-cell line. Eur J Pharmacol 850:88-96 (2019). Read more (PubMed: 30772395) »
  • Chen WC  et al. Xenotropic and polytropic retrovirus receptor 1 (XPR1) promotes progression of tongue squamous cell carcinoma (TSCC) via activation of NF-?B signaling. J Exp Clin Cancer Res 38:167 (2019). Read more (PubMed: 30995931) »
See all 13 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question
Answer


I contacted the laboratories and they confirmed that the active PKA provided in that lot is 430nmol/min/mg and it is at a concentration of 0.02ug/ul.

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Question
Answer



La concentration de la PKA active pour le lot que vous avez reçu (GR140298) est 20 ng/ul. L’activité de la PKA est 498nmol/min/mg.

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