Dead Cell Removal Set, (RUO)

Catalog Number: S0K0009 Brand: Starter

Price:

$500 USD

Size: 1mL

Quantity:

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Product Details

Product Specification

Format	1.Dextran-coated iron oxide magnetic nanoparticles coupled with Annexin V His-tagged protein, in a PBS buffer system containing BSA and Poloxamer 188. 2.Buffer specifically for the dead cell removal kit.
Capacity	0.5 mL magnetic beads: suitable for a total cell count of 5×10⁸and can perform up to 50 sorting runs. 1 mL magnetic beads: suitable for a total cell count of 1×10⁹ and can perform up to 100 sorting runs.
Principle of Separation	The Dead Cell Removal Set can identify a specific structure on the plasma membranes of apoptotic and dead cells. During dead cell removal, cells are first magnetically labeled with Dead Cell Removal Nanobeads, and then the cell suspension is passed through a separation column. The magnetically labeled dead cells are retained within the column, while the unlabeled live cells flow through it; thus, dead cells are removed from this cell fraction. After removing the separation column from the magnetic field, the magnetically retained dead cells can be eluted as the positively selected cell fraction. Using Dead Cell Removal Nanobeads, even early apoptotic cells with intact plasma membranes can be effectively removed.
Applications in cell sorting	Reducing flow cytometry sorting time by removing dead cellsRemoving dead cells from cell culturesImproving Immunocytochemical Analysis of Frozen CellsPreparation of a viable single-cell suspension from tissuesRemoving dead cells from a sperm sampleRemove dead cells before and after cell transfection.
Magnetic Bead Size	20 nm
Separation Method	Column-Based
Selection Strategy	Positive Selection
Stability & Storage	Store protected from light at 2–8°C; do not freeze.

Background

The Dead Cell Removal Set is a rapid and simple method for removing dead cells from cell cultures or tissue preparations. This reagent contains ready-to-use nanomagnetic beads and a binding buffer, which are used to magnetically label cellular debris, dead cells, and dying cells. By magnetic separation, the magnetically labeled material is removed, allowing pure live cells to be obtained in a short time.

Protocol

Using SSeparation Columnas an example

	Steps	Operating Method	Dosage / Time
Magnetic Labeling	1	Count the total number of cells Note:It is recommended to count cells after staining with AO/PI.	—
	2	Centrifuge the cell suspension at 400×g for 7 minutes, and completely discard the supernatant.	400×g, 7 min
	3	Resuspend every 1×10⁷ cells in 90 μL of 1× Binding Buffer. Note:The 20× Binding Buffer must be diluted only with sterile double-distilled water to make 1× Binding Buffer, then add 1% BSA.	Resuspend the cells
	4	Add 10 μL of Dead Cell Removal NanoBeads per 1×10⁷ cells. Note: The binding of Dead Cell Removal NanoBeads requires Ca²⁺. The presence of the ion chelator EDTA will affect binding.	Add 10 μL per 1×10⁷ cells
	5	Mix thoroughly and incubate at 20–25℃ for 15 minutes.	Incubate for 15 minutes
	6	(Optional) If necessary, add 1× binding buffer to the cell suspension to bring the volume to at least 500 μL for separation.	—
Magnetic sorting	7	Place the S-type separation column in the magnetic field of the corresponding magnetic separator, and rinse the column with 500 μL of buffer.	Rinse the separation column
	8	Load the cell suspension onto the separation column and collect the flow-through (live cells).	Collect live cells
	9	Wash the column with buffer three times, 500 μL each time. Add the next wash only after the liquid in the column has completely drained, and combine all the effluents into the negative fraction.	Collect live cells
	10	Remove the separation column from the magnetic field and place it over a collection tube.	—
	11	Add 1 mL of buffer to the column, then quickly and firmly push down with the accompanying plunger to elute the magnetically labeled positive cells. Note:For higher purity, use a new column and repeat the separation once.	Collect the labeled cells

With LSeparation Columnas an example

	Steps	Operating Method	Dosage / Time
Magnetic Labeling	1	Count the total number of cells Note:It is recommended to count cells after AO/PI staining	—
	2	Centrifuge the cell suspension at 400×g for 7 minutes, and completely discard the supernatant	400×g, 7 min
	3	Add 90 μL of 1× Binding Buffer per 1×10⁷ cells to resuspend them Note:The 20× Binding Buffer must be diluted with sterile double-distilled water to 1× Binding Buffer before adding 1% BSA	Resuspend the cells
	4	Add 10 μL of Dead Cell Removal NanoBeads per 1×10⁷ cells Note:The binding of Dead Cell Removal NanoBeads requires Ca²⁺. The presence of the ion chelator EDTA will affect the binding	Add 10 μL per 1×10⁷ cells
	5	Mix thoroughly and incubate at 20–25℃ for 15 minutes	Incubate for 15 minutes
	6	(Optional) If necessary, add 1× binding buffer to the cell suspension to bring the volume to at least 500 μL for separation.	—
Magnetic sorting	7	Place the L-shaped sorting column in the magnetic field of the corresponding magnetic separator, and rinse the column with 3 mL of buffer.	Rinse the sorting column
	8	Load the cell suspension onto the sorting column and collect the flow-through (live cells).	Collect live cells
	9	Wash the column with buffer three times, 3 mL each time. Add the next wash only after the liquid in the column has completely drained. Combine all the effluent as the negative fraction.	Collect live cells
	10	Remove the sorting column from the magnetic field and place it over the collection tube.	—
	11	Add 5 mL of buffer to the column, then quickly and firmly push down with the accompanying plunger to elute the magnetically labeled positive cells. Note:For higher purity, use a new column and repeat the sorting once.	Collect the labeled cells

Picture

Validation Data

Dead cells were removed from C57BL/6 mouse splenocytes using the Dead Cell Removal Set, S or L Separation Columns, and a magnetic sorter. After staining with propidium iodide fluorescence, the samples were analyzed using FlowJo™ software.

Country/region

Dead Cell Removal Set, (RUO)