Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

A novel fluorogenic substrate for detecting alkaline phosphatase activity in situ

Z Huang, W You, R P Haugland

    The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
    |February 1, 1993
    PubMed
    Summary

    A new fluorogenic substrate, CPPCQ, enables sensitive in situ detection of alkaline phosphatase (APase) activity. This method visualizes APase in cells and tissues, offering a novel tool for histochemistry.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    [Analysis of clinical efficacy and perioperative treatment strategies after radical resection for hepatocellular carcinoma with major vascular invasion and tumor thrombus].

    Zhonghua wai ke za zhi [Chinese journal of surgery]·2025
    Same author

    [Surgical treatment and survival analyses of intrahepatic cholangiocarcinoma].

    Zhonghua wai ke za zhi [Chinese journal of surgery]·2025
    Same author

    Relativistic ultrafast electron diffraction at high repetition rates.

    Structural dynamics (Melville, N.Y.)·2023
    Same author

    Reaching adults who smoke cigarettes in rural Appalachia: Rationale, design & analysis plan for a mixed-methods study disseminating pharmacy-delivered cessation treatment.

    Contemporary clinical trials·2023
    Same author

    [Efficacy of non-surgical comprehensive treatment for locally advanced hypopharyngeal carcinoma with cervical esophagus invasion].

    Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery·2023
    Same author

    Pandemic realism as the indispensable political precondition for global disease eradication.

    Public health·2022

    Area of Science:

    • Biochemistry
    • Cell Biology
    • Histochemistry

    Background:

    • Alkaline phosphatase (APase) is an enzyme with diverse biological roles.
    • Accurate detection of APase activity is crucial for various research and diagnostic applications.
    • Existing methods for APase detection may have limitations in sensitivity or specificity.

    Purpose of the Study:

    • To introduce and evaluate a novel fluorogenic substrate, CPPCQ, for the in situ detection of APase activity.
    • To demonstrate the utility of CPPCQ for visualizing both exogenous and endogenous APase.
    • To assess the sensitivity, specificity, and photostability of CPPCQ staining.

    Main Methods:

    • Synthesis and characterization of the fluorogenic substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (CPPCQ).

    Related Experiment Videos

  • Application of CPPCQ for detecting APase activity in fixed human epidermoid carcinoma (A431) cells and fixed canine kidney (MDCK) cells.
  • Utilizing biotinylated EGF and streptavidin-APase conjugates to probe epidermal growth factor (EGF) receptors.
  • Visualization of APase activity and EGF receptors using fluorescence microscopy.
  • Testing specificity by blocking with unlabeled EGF and comparing with fluorescein-labeled EGF.
  • Assessing sensitivity to heat, levamisole, and L-homoarginine to determine APase tissue specificity.

    • Main Results:

    • CPPCQ is a soluble, colorless substrate that yields a fluorescent, precipitating product upon APase cleavage.
    • CPPCQ successfully visualized both biotinylated EGF-anchored APase at EGF receptors and endogenous APase in cell lines.
    • Staining exhibited high contrast, photostability, and resolution under fluorescence microscopy.
    • EGF receptor staining specificity was confirmed by blocking with unlabeled EGF.
    • Endogenous APase staining demonstrated liver/bone/kidney type tissue specificity, consistent with known APase isozymes.

    Conclusions:

    • CPPCQ is a novel and effective fluorogenic substrate for sensitive in situ APase histochemistry.
    • The substrate allows for clear visualization of both targeted and endogenous APase activity.
    • CPPCQ offers advantages in sensitivity, specificity, and imaging quality for APase detection.