Phagocytosis Assay Kit (Zymosan Substrate)

Phagocytosis Assay Kit ab211156 uses a Zymosan substrate to provide a robust system for screening TLR ligands, phagocytosis activators or inhibitors. The phagocytosis assay protocol uses pre-labeled Zymosan particles as a pathogen for triggering phagocytosis.

The engulfed Zymosan particles react with a specific substrate to produce a colorimetric signal that can be detected by absorbance at 405 nm. External particles are blocked before the reaction with a blocking reagent, ensuring the signal is directly proportional to the amount of internalized particles.

This format provides a quantitative, high-throughput method to accurately measure phagocytosis, and avoids subjective manual counting of Zymosan particles inside cells.

Phagocytosis assay protocol summary:
- add Zymosan substrate to cell cultures and incubate for 15 min to 2 hrs
- remove culture medium, and wash cells
- add fixation solution and incubate for 5 min and then wash cells
- add blocking reagent and incubate for 60 min and then wash cells
- add permeabilization solution and incubate for 5 min and then wash cells
- add detection reagent and incubate for 60 min and then wash cells
- add detection buffer and incubate for 10 min
- add substrate and incubate for 5-20 min
- add stop solution and analyze with plate reader

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.

The kit is suitable for adherent phagocytes only.

Each 20 test kit provides sufficient quantities to perform 20, 10 or 5 tests in a 96-,48- or 24-well plate respectively. Each 96 tests kit provides sufficient quantities to perform 96, 48 or 24 tests in a 96-, 48- or 24-well plate respectively. Each 5 x 96 tests kit provides sufficient quantities to perform 5 x 96, 48 or 24 tests in a 96-, 48- or 24-well plate respectively.

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It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.