CD3 Nanobeads, human（RUO）

Catalog Number: S0K0011 Reactivity: Human Conjugation: Brand: Starter

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$285 USD

Size: 1mL 2mL

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Product Details

Product Specification

Format	Dextran-coated iron tetroxide magnetic nanoparticles conjugated with an anti-human CD3 monoclonal antibody.
Capacity	2 mL magnetic beads: suitable for a total cell count of 1×10⁹, and can perform up to 100 sorting runs 1 mL magnetic beads: suitable for 5×10⁸total cells, and can perform up to 50 sorting runs
Principle of Separation	CD3 Nanobeads are used to magnetically label human CD3⁺ cells. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD3⁺ cells remain retained in the column, while the unlabeled cells flow through; this fraction constitutes the negative fraction after removal of CD3⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD3⁺ cells can be eluted, yielding the positive sorted fraction. If higher purity is required, the positive sorted fraction containing CD3⁺ cells must be further separated using a second column.
Reactivity	Human
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

CD3 is expressed on the surface of all T cells and is associated with the T-cell receptor. In human peripheral blood lymphocytes, 70–80% are CD3+, and in thymocytes, 65–85% are CD3+. The antigenic epitope recognized by CD3 Nanobeads is located on the CD3ε chain.

Protocol

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

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	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 CD3 NanoBeads per 1×10⁷ cells Note:When the number of cells increases, increase the volume of all reagents proportionally	Add 20 μL per 1×10⁷ cells
	5	Mix thoroughly, incubate at 2–8℃ in the dark for 15 minutes	Incubate for 15 min
	6	Add 1 mL of buffer to wash every 1×10⁷ cells, centrifuge at 400×g for 7 minutes, and completely discard the supernatant	Add 1 mL to 1×10⁷ cells
	7	Resuspend up to 1×10 in 500 μL of buffer⁸Cells	Resuspend the cells
Magnetic separation	8	Place the S-shaped 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 effluent (unlabeled cells).	—
	10	Wash the column with buffer three times, 500 μL each time, adding the next wash only after the liquid in the column has completely drained.	3 × 500 μL of buffer
	11	Combine all effluent fractions into the negative fraction; remove the separation column from the magnetic field and place it on the collection tube.	—
	12	Add 1 mL of buffer to the column, and quickly and firmly push 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 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 CD3 NanoBeads per 1 × 10⁷ cells. Note:When the number of cells increases, increase the volume of all reagents proportionally.	Add 20 μL per 1 × 10⁷ cells.
	5	Mix thoroughly, incubate at 2–8℃ in the dark for 15 min	Incubate for 15 min
	6	Add 1 mL of buffer to wash every 1×10⁷ cells, centrifuge at 400×g for 7 minutes, and completely discard the supernatant	1×10⁷ cells plus 1 mL
	7	Resuspend up to 1×10 in 500 μL of buffer⁸Cells	Resuspend 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 effluent (unlabeled cells)	—
	10	Wash the column three times with 3 mL of buffer each time, adding the next rinse only after the liquid inside the column has completely drained	3 × 3 mL of buffer
	11	Combine all effluents 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

Enrich or deplete T cells from human PBMCs. Cells were stained with anti-CD3 (clone: UCHT1) Brilliant Violet 421™, and the live cell population was gated for analysis.

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CD3 Nanobeads, human（RUO）