CytoSelect™ 96-well Phagocytosis Assay (Red Blood Cell)

Product Description

• Fast and accurate quantification of phagocytosis
• Protocol based on engulfing red blood cells
• No manual counting
• High-throughput compatible

Traditionally, erythrocytes (red blood cells) are commonly used in phagocytosis assays. For FcR mediated phagocytosis, erythrocytes are first opsonized with serum or IgG before they are added to phagocytes. After removal of non-phagocytic erythrocytes, engulfed erythrocytes are quantitated.

In contrary to conventional phagocytosis assays, the CytoSelect™ 96-well Phagocytosis Assay does not involve subjective manual counting of erythrocytes. Instead cells are lyzed and detected by the proprietary erythrocyte substrate in a microtiter plate reader. This format provides a quantitative, high-throughput method to accurately measure phagocytosis.

Each kit provides sufficient quantities to perform 96, 48, 24 tests in a 96, 48, 24-well plate, respectively.

Assay Principle

Fig.1. Principle of the CytoSelect™ 96-well Phagocytosis Assay.

Performance Data

Fig.2. Inhibition of Raw 264.7 macrophage phagocytosis by cytochalasin D.
20,000 cells/well of Raw 264.7 macrophages were seeded overnight in a 96-well plate. Cytochalasin D was used to pre-treat Raw 264.7 cells for 1 hr at 37 ºC before IgG opsonized sheep red blood cells (RBC) were added at 50:1 ratio. Phagocytosis was stopped after 30 minutes and the amount of engulfed red blood cells was determined as described in the protocol of the CytoSelect™ 96-well Phagocytosis Assay (Erythrocytes based). (A) Non-opsonized RBC + Raw 264.7. (B) IgG opsonized RBC + Raw 264.7. (C) IgG opsonized RBC + Raw 264.7 (2 µM cytochalasin D pre-treated).