Phagocytosis Assay Kit (Zymosan, Colorimetric)
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(8 Publications)
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Supplier Data
Functional Studies - Phagocytosis Assay Kit (Zymosan, Colorimetric) (AB211156)
Zymosan Particles Engulfment by Raw 264.7 Macrophage
- FuncS
Supplier Data
Functional Studies - Phagocytosis Assay Kit (Zymosan, Colorimetric) (AB211156)
Inhibition of Raw 264.7 Macrophage Phagocytosis by Cytochalasin D.
50,000 cells/well of Raw 264.7 macrophages were seeded overnight in a 96-well plate. Cytochalasin D was used to pretreat Raw 264.7 cells for 1 hr at 37°C before addition of Zymosan at a ratio of 50 : 1 Zymosan particles per cell. Phagocytosis was stopped after 30 minutes and the amount of engulfed Zymosan particles was determined as described in the Assay Protocol.
Product details
Phagocytosis Assay Kit ab211156 uses a Zymosan substrate to provide a robust system for screening TLR ligands, phagocytosis activators or inhibitors.
Colorimetric phagocytosis assay principle
The phagocytosis assay uses pre-labeled Zymosan particles as a pathogen to trigger phagocytosis. The label on the engulfed Zymosan particles then reacts with a 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
Getting the best performance from ab211156
This 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.
Related phagocytosis assays
Phagocytosis Assay Kit (Green E.coli) ab235900
Phagocytosis Assay Kit (Red E. coli) ab235901
Phagocytosis Assay Kit (Green Zymosan) ab234053
Phagocytosis Assay Kit (Red Zymosan) ab234054
Other 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.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
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.
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Publications (8)
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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