Overview

  • Product name

    Phagocytosis Assay Kit (Red E. coli)
    See all Phagocytosis kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Product overview

    Phagocytosis Assay Kit (Red E. coli) (ab235901) utilizes heat-killed, fluorescently pre-labeled E. coli particles as a tool for rapid and accurate detection and quantification of in vitro phagocytosis by fluorescent microscope, spectrophotometer or flow cytometry. The kit provides a robust screening system for activators and/or inhibitors of phagocytosis and Toll-like receptor (TLR) ligands.

  • Notes

    Phagocytosis in mammals serves as an important first line defense mechanism against invading pathogens. It is also essential for continuous clearance of dying cells, tissue remodeling, and acquisition of nutrients for some cells. Phagocytosis is a specific form of endocytosis initiated by recognition and binding of foreign particles by cell surface receptors, followed by their engulfment, and formation of phagosomes. Maturing phagosomes transform to phagolysosomes which destroy the pathogen through enzymes and toxic peroxides. E. coli and other bacterial strains are often used as a pathogen in phagocytosis assays.

  • Platform

    Microplate reader, Fluor. microscope, Flow cyt.

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    10X Quenching Solution 1 x 500µl
    Buffer Additive 2 x 1ml
    Phagocytosis Assay Buffer  2 x 100ml
    Red E. coli 1 x 600µl

Images

  • Inhibition of phagocytosis. 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 hour at 37ºC prior to addition of 5 µL of E. coli particles. Phagocytosis was conducted for 2 hours and the amount of engulfed E. coli was determined as described in the Assay Protocol. Panel A and B: images of nontreated 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 hour at 37ºC prior to addition of 5 µL of E. coli particles. Phagocytosis was conducted for 2 hours and the amount of engulfed E. coli was determined as described in the Assay Protocol. Blue line: untreated control cells; red line: macrophages with engulfed E. coli particles; green line: inhibition of phagocytosis by Cytochalasin D.

  • E. coli Standard curve.

Protocols

References

ab235901 has not yet been referenced specifically in any publications.

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