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

In vitro bioassay for transforming growth factor-beta using XTT method.

Mi-Sung Kim1, Seong-Min Ahn, Aree Moon

  • 1College of Pharmacy, Duksung Womens University, Seoul 132-714, Korea.

Archives of Pharmacal Research
|January 4, 2003
PubMed
Summary

This study introduces a safer and more sensitive XTT assay for measuring transforming growth factor-beta (TGF-beta) activity in vitro. The optimized XTT assay offers a lower detection limit than traditional methods, improving cytokine research.

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

Recent Advances in Donepezil Delivery Systems via the Nose-to-Brain Pathway.

Pharmaceutics·2025
Same author

Cell enlargement modulated by GATA4 and YAP instructs the senescence-associated secretory phenotype.

Nature communications·2025
Same author

Regulation of senescence-associated secretory phenotypes in osteoarthritis by cytosolic UDP-GlcNAc retention and O-GlcNAcylation.

Nature communications·2025
Same author

Autophagy-dependent splicing control directs translation toward inflammation during senescence.

Developmental cell·2024
Same author

Metabolic remodeling in cancer and senescence and its therapeutic implications.

Trends in endocrinology and metabolism: TEM·2024
Same author

ANXA2 (annexin A2) is crucial to ATG7-mediated autophagy, leading to tumor aggressiveness in triple-negative breast cancer cells.

Autophagy·2024

Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Cytokine research is rapidly expanding, necessitating precise measurement techniques.
  • Transforming growth factor-beta (TGF-beta) is a key regulator of cell growth and differentiation.
  • Current TGF-beta bioassays, like the mink lung cell proliferation assay using [3H]thymidine incorporation or MTT assay, have limitations regarding sensitivity, safety, and disposal.

Purpose of the Study:

  • To develop a safer and more sensitive in vitro method for quantifying TGF-beta biological activity.
  • To compare the XTT assay with the conventional MTT assay under various experimental conditions.

Main Methods:

  • Compared MTT and XTT assays for TGF-beta detection.
  • Optimized parameters including cell number, incubation time, and wavelength.

Related Experiment Videos

  • Tested anti-proliferative activity of TGF-beta on Mv-1-Lu, MCF10A, and H-ras-transformed MCF10A cells.

    • Main Results:

    • The optimized XTT assay demonstrated a detection limit as low as 10 pg/ml for TGF-beta.
    • Identified optimal conditions: Mv-1-Lu or H-ras MCF10A cells (1 x 10(5)/well) incubated with TGF-beta for 24 hr, followed by XTT treatment and absorbance reading at 450 or 490 nm.
    • The XTT assay proved more sensitive and safer than radioisotope-based methods.

    Conclusions:

    • An optimized XTT assay protocol provides a highly sensitive and safe method for quantifying in vitro TGF-beta biological activity.
    • This improved bioassay system can facilitate satisfactory quantitation of TGF-beta.
    • The findings contribute to advancing cytokine measurement techniques in biological research.