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Transglutaminase Activity Assay Kit (Colorimetric) (ab204700)

  • Datasheet
  • SDS
  • Protocol Booklet
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Transglutaminase Activity Assay Kit (ab204700)
  • Transglutaminase Activity Assay Kit (ab204700)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 2 hr 30 min
  • Sample type: Adherent cells, Suspension cells, Tissue
  • Sensitivity: 10 µU

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Overview

  • Product name

    Transglutaminase Activity Assay Kit (Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    Tissue, Adherent cells, Suspension cells
  • Assay type

    Enzyme activity
  • Sensitivity

    10 µU
  • Assay time

    2h 30m
  • Product overview

    Transglutaminase Activity Assay Kit (ab204700) utilizes the deamidation reaction of the transglutaminase enzyme with a donor and acceptor substrate resulting in the formation of a hydroxamate product. The hydroxamate product reacts with the Stop Solution forming a purple complex that can be measured colorimetrically at 525 nm. The limit of quantification of this assay is ~10 µU in cell/tissue lysates.


    Transglutaminase assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 2 hrs
    - add stop solution and centrifuge plate at 1800 g for 15 min
    - transfer supernatant to fresh plate
    - analyze with microplate reader

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K571 Transglutaminase Activity Assay Kit (Colorimetric). K571-100 is the same size as the 100 test size of ab204700.

    Transglutaminases (EC 2.3.2.13) are calcium dependent enzymes that catalyze the post-translational modification of proteins by formation of isopeptide bonds. This occurs either through protein cross-linking via formation of γ-glutamyl-ε-lysine bonds or through incorporation of primary amines at selected peptide-bound glutamine residues. The transglutaminase enzyme family comprises the intracellular forms (TG1, TG3 and TG5) expressed mostly in the epithelial tissue; TG2 which is both intracellular and extracellular and expressed in various tissue types; TG4 which is expressed in the prostate gland; factor XIII which is expressed in blood; TG6 and TG7, whose tissue distribution is unknown and band 4.2 (lacking enzymatic activity) which is present on erythrocyte membranes. Transglutaminases also exhibit GTPase, phosphodiesterase and protein kinase activity. Transglutaminases are associated with certain neurological and autoimmune disorders and also cancer.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Acceptor Substrate 2 vials
    Donor Substrate 1 vial
    DTT 1 x 125µl
    Homogenization Buffer (10x) 1 x 10ml
    Hydroxamate Standard 1 vial
    Plate Sealer 1 unit
    Positive Control 1 vial
    Stop Solution 1 x 5ml
    TG Assay Buffer 1 x 12ml
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calcium Binding Proteins
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
  • Relevance

    Transglutaminases (EC 2.3.2.13) are calcium dependent enzymes that catalyze the post-translational modification of proteins by formation of isopeptide bonds. This occurs either through protein cross-linking via formation of ?-glutamyl-e-lysine bonds or through incorporation of primary amines at selected peptide-bound glutamine residues. The transglutaminase enzyme family comprises the intracellular forms (TG1, TG3 and TG5) expressed mostly in the epithelial tissue; TG2 which is both intracellular and extracellular and expressed in various tissue types; TG4 which is expressed in the prostate gland; factor XIII which is expressed in blood; TG6 and TG7, whose tissue distribution is unknown and band 4.2 (lacking enzymatic activity) which is present on erythrocyte membranes. Transglutaminases also exhibit GTPase, phosphodiesterase and protein kinase activity. Transglutaminases are associated with certain neurological and autoimmune disorders and also cancer.

Images

  • Transglutaminase Activity Assay Kit (ab204700)
    Transglutaminase Activity Assay Kit (ab204700)

    Typical Hydroxamate Standard calibration curve.

  • Transglutaminase Activity Assay Kit (ab204700)
    Transglutaminase Activity Assay Kit (ab204700)

    Transglutaminase activity in HepG2 cells (human hepatoblastoma cell line) and rat liver lysate: HepG2 cells were stimulated with vehicle (DMSO), IL6 (1 µM), Dexamethasone (DXM -1 µM), or with IL6 (1µM) and DXM (1 µM). Approximately 250 µg protein was used for determining transglutaminase activity in cells and tissue lysate. Activity is expressed as nmoles of product formed in 2h and is normalized to the protein amount.

    Note: HepG2 cells have similar intrinsic Transglutaminase activity in the presence or absence of vehicle control (DMSO).

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab204700? Please let us know so that we can cite the reference in this datasheet.

ab204700 has been referenced in 2 publications.

  • Caldwell CC  et al. Treadmill exercise rescues mitochondrial function and motor behavior in the CAG140 knock-in mouse model of Huntington's disease. Chem Biol Interact 315:108907 (2019). PubMed: 31778667
  • Monguzzi E  et al. Gliadin effect on the oxidative balance and DNA damage: An in-vitro, ex-vivo study. Dig Liver Dis N/A:N/A (2018). PubMed: 30055963

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