Overview

  • Product name

    Phagocytosis Assay Kit (Red Zymosan)
    See all Phagocytosis kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Phagocytosis Assay Kit ab234054 uses Zymosan particles which have been pre-labeled with a red dye.


    The assay can be used for the rapid and accurate detection and quantification of in vitro phagocytosis by fluorescent microscope, spectrophotometer or flow cytometry. It rovides a robust screening system for activators and/or inhibitors of phagocytosis and Toll-like receptors ligands (TLR).


    The Zymosan particles used in the phagocytosis assay protocol are labeled for fluorescence at Ex/Em 540/570 nm.


    Phagocytosis assay protocol summary:
    - add Zymosan to cells and incubate for 2-3 hrs
    - harvest cells and resuspend pellet in phagocytosis assay buffer, repeat three times to wash off any remaining Zymosan which is not inside cells
    - analyze cells with flow cytometer, fluorescent microscope, or microplate reader

  • Notes

    In mammals, phagocytosis by phagocytes (e.g., macrophages, dendritic cells, and neutrophils) is essential for a variety of biological events. Phagocytosis comprises a series of events, starting with the binding and recognition of particles by cell surface receptors, followed by the formation of actin-rich membrane extensions around the particle.  

    Zymosan (Saccharomyces cerevisiae) is prepared from yeast cell wall and consists of protein-carbohydrate complexes.  Zymosan is a commonly used pathogen in phagocytosis assays.

  • Platform

    Microplate reader, Fluor. microscope, Flow cyt.

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 100 tests
    Phagocytosis Assay Buffer 2 x 100ml
    Buffer Additive 2 x 1ml
    Red Zymosan (Cy3) 1 x 600µl
    10X Quenching Solution 1 x 500µl

Images

  • J774 macrophages were seeded overnight at 5 x 105 of viable cells/well. The next day the cells were pretreated with 20 µM Cytochalasin D for 1 h at 37ºC prior to addition of 5 µl of Zymosan particles. Phagocytosis was conducted for 2 hours and the amount of engulfed Zymosan was determined as described in the Assay Protocol.

    Panel A and B: images of non-treated cells. Panel C and D: treatment with Cytochalasin D.

  • J774 macrophages were seeded overnight at 5 x 105 of viable cells/well. The next day the cells were pretreated with 20 µM Cytochalasin D for 1 h at 37ºC prior to addition of 5 µl of Zymosan particles. Phagocytosis was conducted for 2 hours and the amount of engulfed Zymosan was determined as described in the Assay Protocol.

    Black line: untreated control cells; red line: macrophages with engulfed Zymosan particles; blue line: inhibition of phagocytosis by Cytochalasin D.

  • J774 macrophages were seeded overnight at 5 x 105 of viable cells/well. The next day the cells were pretreated with 20 µM Cytochalasin D for 1 h at 37ºC prior to addition of 5 µl of Zymosan particles. Phagocytosis was conducted for 2 hours and the amount of engulfed Zymosan was determined as described in the Assay Protocol.

Protocols

References

ab234054 has not yet been referenced specifically in any publications.

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