CD4 Nanobeads, mouse（RUO）

Catalog Number: S0K0003 Reactivity: Mouse Conjugation: Brand: Starter

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$283 USD

Size: 1ml 2ml

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Product Details

Product Specification

Format	Dextran-coated iron tetroxide nanomagnetic beads conjugated with an anti-mouse CD4 monoclonal antibody
Capacity	2 mL magnetic beads: suitable for a total cell count of 1×10⁹, and can perform up to 100 separations 1 mL magnetic beads: suitable for 5×10⁸total cells, and can perform up to 50 separations
Principle of Separation	CD4⁺ cells are magnetically labeled using CD4 Nanobeads, mouse. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD4⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD4⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD4⁺ cells can be eluted, yielding the positive sorted fraction. To further enhance purity, the positive fraction containing CD4⁺ cells must be subjected to 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

CD4 antigen is expressed on most thymocytes and mature helper T cells, certain NKT cell subsets, and a small fraction of dendritic cell subsets. In CD4⁺helper T cells, CD4 acts as a co-molecule to assist the TCR heterodimer in recognizing MHC class II/peptide complexes. CD4 antigen is present on approximately 90% of thymocytes, 25% of splenic cells, and 55% of lymph node cells.

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 80 μL of buffer to resuspend every 1×10⁷ cells Note:Adjust the volume of the buffer proportionally according to the sample volume	Resuspend the cells
	4	Add 20 μL of CD4 NanoBeads per 1×10⁷ cells Note:When the cell quantity increases, increase the volume of all reagents proportionally	Add 20 μL per 1×10⁷ cells
	5	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes	Incubate for 15 minutes
	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 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 80 μL of buffer. Note:Adjust the volume of the buffer proportionally according to the sample volume.	Resuspend the cells.
	4	Add 20 μL of CD4 NanoBeads per 1×10⁷ cells. Note:When the number of cells increases, increase the volumes of all reagents proportionally at the same time.	Add 20 μL per 1×10⁷ cells.
	5	Mix thoroughly and incubate in the dark at 2–8°C 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 separation	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 with buffer three times, 3 mL each time, adding the next wash only after the liquid in 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

CD4+ T cells were enriched from or depleted of spleen cells isolated from BALB/c mice. Subsequently, fluorescent staining was performed using anti-CD4 (clone: GK1.5) Brilliant Violet 421™, and the live cell population was gated for analysis.

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CD4 Nanobeads, mouse（RUO）