CD49b Nanobeads, mouse（RUO）

Catalog Number: S0K0017 Reactivity: Mouse Conjugation: Brand: Starter

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$500 USD

Size: 1mL 2mL

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Product Details

Product Specification

Format	Dextran-coated iron tetroxide magnetic nanoparticles conjugated with an anti-mouse CD49b monoclonal antibody
Capacity	2 mL magnetic beads: suitable for a total cell count of 2 × 10⁹, allowing up to 200 sorting runs 1 mL magnetic beads: suitable for a total cell count of 1 × 10⁹, allowing up to 100 sorting runs
Principle of Separation	AdoptCD49b Nanobeads, mousefor magnetic labeling of CD49b cells. Subsequently, the cell suspension is loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD49b cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD49b cells. After removing the separation column from the magnetic field, the magnetically captured CD49 cells can be eluted, yielding the positive sorted fraction. If higher purity is required, the positive sorted fraction containing CD49b cells should be further separated using 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

CD49b Nanobeads, mouse are designed to isolate mouse NK cells from single-cell suspensions of lymphoid and non-lymphoid tissues, peripheral blood, or body fluids. CD49b is expressed on NK cells and a small subset of T cells (CD4⁺CD3⁺TCRαβ⁺). Compared with other NK cell markers such as NK1.1, CD49b exhibits lower strain specificity in mice and is expressed in most commonly used inbred mouse strains.

Protocol

Note: This kit does not include separation 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 volume of buffer proportionally according to the sample volume	Resuspend the cells
	4	Add 10 μL of CD49b Nanobeads per 1×10⁷ cells Note:When the cell quantity increases, increase the volume of all reagents 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 discard the supernatant.	Add 1 mL to 1×10⁷ cells
	7	Resuspend up to 1×10⁸ cells in 500 μL of buffer⁸Cells	Resuspend 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 CD49b Nanobeads per 1×10⁷ cells. Note:When the number of cells increases, increase the volumes of all reagents proportionally.	Add 10 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 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 with buffer three times, 3 mL each time, and add the next wash 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

CD49b⁺ cells were isolated from the single-cell suspension using CD49b Nanobeads, S or L Separation Columns, and a magnetic separator. Subsequently, fluorescent staining was performed with Alexa Fluor® 488 Armenian hamster Anti-Mouse CD3 Antibody (S-R491) (STD Cat: S0B8364) and Alexa Fluor® 647 Rat Anti-Mouse CD49b Antibody (DX5) (STD Cat: S0B8257), and the live cell population was gated for analysis.

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CD49b Nanobeads, mouse（RUO）