Lysosomal Intracellular Activity Assay Kit (ab234622)

Overview

  • Product name

    Lysosomal Intracellular Activity Assay Kit
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Lysosomal Intracellular Activity Assay Kit (ab234622) provides a proprietary Lysosome-Specific Self-Quenched Substrate which has low background fluorescence, high signal to background ratio and is pH insensitive. The substrate, acting as endocytic cargo, can be taken up by cells and degraded within an endo-lysosomal vesicle. The fluorescent signal is recovered from the Self-Quenched Substrate. The fluorescence signal, generated by degradation, is proportional to the intracellular lysosomal activity and can be measured using a fluorescence microscopy and/or flow cytometry. The kit includes Cytochalasin D, a cell-permeable inhibitor of endocytosis that serves as an experimental control. This easy-to-use, non-radioactive kit allows imaging and accurate measurement of de-quenching substrate in cultured cells.

  • Notes

    Lysosomes are membrane-bound organelles important for various cellular processes. They contain hydrolytic enzymes that are utilized in the metabolism of some biomolecules. The extracellular cargo (e.g. nutrients toxins) binds to the cell membrane and is subsequently transported into membrane-bound endosomes for further degradation by lysosomes while intracellular components are transported to lysosomes through autophagy. Lysosomal dysfunction is associated with many human conditions such as aging and neurodegenerative disease.

  • Platform

    Flow cytometer, Fluorescence microscope

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 50 tests
    Assay Buffer (50X) 1 x 1.8ml
    Self-Quenched Substrate 1 vial
    Cytochalasin D (100X) 1 x 50µl

Images

  • 1x106 U937 cells were pretreated with vehicle or 1X Cytochalasin D for 1 hour. After pre-treatment, cells were incubated with Self-Quenched Substrate and the same concentration of Cytochalasin D for additional hour in medium supplemented with 0.5% FBS according to kit’s protocol. Comparison of histograms from flow analysis showing the inhibition of De-Quenching of Substrate by Cytochalasin D. Unstained cells (black); experimental control (green) in the presence of 1X Cytochalasin D; Positive control (pink) without 1X Cytochalasin D.

  • 1x106 U937 cells were pretreated with vehicle or 1X Cytochalasin D for 1 hour. After pre-treatment, cells were incubated with Self-Quenched Substrate and the same concentration of Cytochalasin D for additional hour in medium supplemented with 0.5% FBS according to kit’s protocol. Images of U937 cells obtained using fluorescence microscope. Top: positive control cells treated Self-quenched substrate only. Bottom: negative control cells treated with 1X Cytochalasin D. U937 cells showing the release of Self-quenched substrate in the endocytotic vesicle (punctured pattern).

  • 1x106 U937 cells were pretreated with vehicle or 1X Cytochalasin D for 1 hour. After pre-treatment, cells were incubated with Self-Quenched Substrate and the same concentration of Cytochalasin D for additional hour in medium supplemented with 0.5% FBS according to kit’s protocol. Lysosomal staining with Lysosomal Associated Membrane Protein 1 (Lamp-1 RFP, a lysosomal marker). Cells were stained with nuclear dye for 10 minutes, washed with 1X PBS and mounted on the slide. Images were taken using a fluorescence microscope with a 60X objective lens.

Protocols

References

ab234622 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab234622.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up