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Cat. No.: DCRKV-100 (for 100T)

Cat. No.: DCRKV-400 (for 400T)

Description

Dead Cell Removal Kit (Annexin V)—also referred to as the Apoptotic Cell Removal Kit (Annexin V) or Live Cell Enrichment Kit—is a magnetic bead–based system that utilizes Annexin V recombinant protein in combination with antibody‑conjugated magnetic beads to rapidly, efficiently, and conveniently remove apoptotic and other dead cells from cell samples within approximately 30 minutes. Cells processed with this kit exhibit high viability and can be further cultured or directly used for downstream applications such as functional assays, gene expression analysis, and phenotypic characterization. The kit is generally used for negative selection, but it can also serve as a positive selection tool for isolating and enriching apoptotic or dead cells.

The selection and removal performance of this kit is comparable to products such as STEMCELL Technologies’ EasySep™ Dead Cell Removal (Annexin V) Kit and Miltenyi Biotec’s Dead Cell Removal Kit. It does not require specialized separation columns or magnetic separators—only a standard magnetic rack is needed.

Annexins are a family of calcium‑dependent phospholipid‑binding proteins widely distributed in the cytoplasm of eukaryotic cells and involved in intracellular signal transduction. Among them, only Annexin V has been reported to regulate certain PKC activities. Annexin V selectively binds to phosphatidylserine (PS), which is normally located on the inner leaflet of the plasma membrane, adjacent to the cytoplasm. During early apoptosis, many cell types translocate PS to the outer leaflet of the membrane. Once exposed on the cell surface, PS promotes coagulation and inflammatory responses. Binding of Annexin V to externalized PS blocks these pro‑coagulant and pro‑inflammatory activities. The Annexin V recombinant protein used in this kit is labeled with EGFP, enabling direct visualization and assessment of apoptotic/dead cell removal using flow cytometry or fluorescence microscopy.

The working principle of the kit is as follows: Annexin V‑EGFP binds to PS exposed on apoptotic/dead cells, which then interact with antibody‑conjugated magnetic beads (Binding Magnetic Beads). After magnetic separation using a magnetic rack or similar device, apoptotic cells are captured by the beads, and the resulting supernatant contains a highly viable cell population with dead cells removed.

This kit demonstrates excellent performance in removing apoptotic and dead cells, achieving a removal efficiency of 90–95%, outperforming comparable products.

The kit is simple and fast to use. Based on a magnetic bead–based workflow, it does not require chromatography columns or complicated separation procedures. Dead cells can be easily and rapidly removed using a standard magnetic rack, and the entire process can be completed in approximately 30 minutes.

The kit offers broad applicability and flexible detection options. It is compatible with most cell types and does not require species differentiation, making it suitable for virtually all eukaryotic cells that contain phosphatidylserine. The efficiency of dead cell removal can be verified using either flow cytometry or fluorescence microscopy.

Assuming 5 million cells per experiment, the small‑size and medium‑size packages of this kit can perform approximately 100 and 400 dead‑cell removal procedures, respectively, when the proportion of apoptotic/dead cells in the sample is below 30%. For samples in which apoptotic/dead cells exceed 50%, a higher amount of reagents is required to achieve optimal removal efficiency; under these conditions, the small‑size and medium‑size packages can be used for approximately 50 and 200 procedures, respectively. For samples with less than 10% apoptotic/dead cells, reagent volumes can be reduced by half while still achieving ideal removal performance, allowing the small‑size and medium‑size packages to be used for a greater number of procedures.

Components

• Annexin V-EGFP
• Binding Magnetic Beads
• Dead Cell Removal Buffer

Storage

Store at –20ºC for up to one year; store at 4ºC for at least three months. Annexin V‑EGFP must be protected from light during storage.

Precautions

• If the processed cells will be used for subsequent culture, prepare 70% ethanol in advance for sterilizing the Binding Magnetic Beads.
• Although testing shows that Annexin V‑EGFP retains its performance after five freeze–thaw cycles, for optimal results it is recommended to store the reagent at 4ºC protected from light and avoid repeated freeze–thawing when used within three months.
• Bacterial or fungal contamination will severely compromise experimental results.
• If trypsin was used during cell harvesting, ensure that residual trypsin is removed. Remaining trypsin can digest and degrade Annexin V‑EGFP, ultimately reducing dead‑cell removal efficiency.
• The Dead Cell Removal Buffer in this kit contains a certain concentration of Ca²⁺. Before starting the experiment, ensure that the cell resuspension buffer does not contain EDTA or contains only a very low level of EDTA, otherwise the removal efficiency may be affected.
• Do not leave the Binding Magnetic Beads in a magnetic field for extended periods, as this may cause bead aggregation.
• Before use, the Binding Magnetic Beads should be fully resuspended by gently inverting the tube several times to ensure uniform mixing. Avoid vigorous vortexing or shaking to prevent antibody denaturation.
• This product is intended for research use only by trained professionals. It is not for clinical diagnosis or therapy, not for use in food or pharmaceuticals, and must not be stored in residential settings.
• For your safety and health, wear a lab coat and disposable gloves during operation.

Only for research and not intended for treatment of humans or animals

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