CD8 Nanobeads, mouse（RUO）

Catalog Number: S0K0005 Reactivity: Mouse Conjugation: Brand: Starter

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$533 USD

Size: 1ml 2mL

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Product Details

Product Specification

Format	Dextran-coated iron tetroxide magnetic nanoparticles conjugated with anti-mouse CD8 monoclonal antibody
Capacity	2 mL magnetic beads: suitable for a total cell count of 2 × 10⁹, and can perform up to 200 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	CD8⁺ cells are magnetically labeled using CD8 Nanobeads, mouse. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD8⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD8⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD8⁺ cells can be eluted, yielding the positive sorted fraction. If higher purity is required, the positive fraction containing CD8⁺ cells must be further separated through a second separation column.
Reactivity	Mouse
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

CD8 NanobeadsThese beads are designed for positive selection or depletion of murine cytotoxic T cells from lymphoid organs, blood, or tissues. The magnetic beads react with the immunoglobulin-like domain of the CD8α chain. The CD8α chain is expressed as a heterodimer with CD8β on cytotoxic T cells that express either the T cell receptor (TCR) α/β or TCR γ/δ, as well as on most thymocytes. As a homodimer composed of two CD8α chains, the CD8 antigen is present on most NK cells, the majority of intraepithelial lymphocytes in the intestine, some activated CD4+CD8+ T cells, and on CD8+ T cells in thymus-deficient mice.

Protocol

Note: This kit does not include the sorting buffer; please prepare it yourself. The corresponding Stater catalog number isS0D3018

Taking S as an exampleSeparation ColumnFor instance

	Steps	Operating procedures	Dosage / Time
Magnetic labeling	1	Count the total number of cells	—
	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 buffer to resuspend every 1×10⁷ cells Note:Adjust the amount of buffer proportionally based on the sample volume	Resuspend the cells
	4	Add 10 μL of CD8 NanoBeads per 1×10⁷ cells Note:When the cell quantity increases, all reagent volumes should be increased proportionally	Add 10 μL per 1×10⁷ cells
	5	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes	Incubate for 15 minutes
	6	Add 1 mL of buffer to wash every 1×10⁷ cells, centrifuge at 400×g for 7 minutes, and completely aspirate the supernatant.	Add 1 mL to 1×10⁷ cells
	7	Resuspend up to 5×10⁷ cells in 500 μL of buffer	Resuspend the cells
Magnetic separation	8	Place the S-type separation column into the magnetic field of the corresponding magnetic separator, and rinse the column with 500 μL of buffer.	Rinse the separation column
	9	Load the cell suspension onto the separation column and collect the flow-through (unlabeled cells).	—
	10	Wash the column three times with 500 μL of buffer each time, adding the next wash only after the liquid in the column has completely drained.	3 × 500 μL of buffer
	11	Combine all flow-through fractions into the negative fraction; remove the separation column from the magnetic field and place it over the collection tube.	—
	12	Add 1 mL of buffer to the column, and quickly and firmly push 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 target cells.

With LSeparation Columnas an example

	Steps	Operating method	Dosage / Time
Magnetic labeling	1	Count the total number of cells	—
	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 buffer. Note:Adjust the volume of the buffer proportionally according to the sample volume.	Resuspend the cells.
	4	Add 10 μL of CD8 NanoBeads per 1×10⁷ cells. Note:When the cell quantity increases, increase the volumes of all reagents proportionally at the same time.	Add 10 μL per 1×10⁷ cells.
	5	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes.	Incubate for 15 min.
	6	Wash every 1×10⁷ cells with 1 mL of buffer, centrifuge at 400×g for 7 minutes, and completely discard the supernatant.	Add 1 mL to 1×10⁷ cells
	7	Resuspend up to 5×10⁷ cells in 500 μL of buffer	Resuspend the cells
Magnetic sorting	8	Place the L-shaped separation column into the magnetic field of the corresponding magnetic separator, and rinse the column with 3 mL of buffer.	Rinse the separation column
	9	Load the cell suspension onto the separation column and collect the flow-through (unlabeled cells).	—
	10	Wash the column three times with 3 mL of buffer each time, adding the next wash only after the liquid inside the column has completely drained.	3 × 3 mL of buffer
	11	Combine all flow-through fractions into the negative fraction; remove the separation column from the magnetic field and place it over the collection tube.	—
	12	Add 5 mL of buffer to the column, and use the accompanying plunger to quickly and firmly push, eluting the magnetically labeled positive cells. Note:For higher purity, repeat the separation once using a new column.	Collect the target cells.

Picture

Validation Data

CD8+ T cells were enriched from or depleted of splenocytes isolated from C57BL/6 mice. Subsequently, fluorescent staining was performed using anti-CD8 (clone: 53-6.7) Alexa Fluor® 647, and the live cell population was gated for analysis.

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CD8 Nanobeads, mouse（RUO）