CD11c Nanobeads Kit, mouse (RUO)

Catalog Number: S0K0022 Reactivity: Mouse Conjugation: Brand: Starter

Price:

$500 USD

Size: 1 mL 2 mL

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Product Details

Product Specification

Format	Dextran-coated iron oxide nanomagnetic beads conjugated with an anti-mouse CD11c monoclonal antibody, in a PBS buffer system containing BSA and Poloxamer 188.
Capacity	1 mL magnetic beads: suitable for 1×10^nineTotal cell count: up to 100 sorting runs can be performed. 2 mL magnetic beads: suitable for a total cell count of 2 × 10⁹, enabling up to 200 sorting runs.
Principle of Separation	CD11c⁺ cells are magnetically labeled using CD11c Nanobeads, mouse. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD11c⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after depletion of CD11c⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD11c⁺ cells can be eluted, yielding the positive sorted fraction.
Applications in cell sorting	Isolate or deplete CD11c+ cells from a single-cell suspension of mouse spleen.
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

CD11c Nanobeads are specifically designed for the isolation of CD11c-positive cells (such as dendritic cells, DCs) from single-cell suspensions derived from murine lymphoid and non-lymphoid tissues. Unlike human cells, all known subsets of dendritic cells in mice express CD11c, and certain tissue-resident macrophage subsets also carry this marker. By using these CD11c magnetic beads, you can replace the traditionally laborious and complex steps of dendritic cell isolation with a simple and rapid positive-selection protocol.

Protocol

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

Taking S Separation Column as an example

	Steps	Operating method	Dosage / Time
Magnetic labeling	1	Take the mouse spleen, grind it on a 40 μm sterile filter to prepare a single-cell suspension of the spleen; after red blood cell lysis, proceed with cell counting 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 buffer. Note:Adjust the volume of the buffer proportionally according to the sample volume.	Resuspend the cells
	4	Add 10 μL of CD11c Nanobeads per 1×10⁷ cells Note:When the cell quantity increases, increase the volume of all reagents proportionally at the same time.	1 × 10⁷ cells plus 10 μL
	5	Mix thoroughly, incubate in the dark at 2–8°C for 15 minutes	Incubate for 15 min
	6	Add 1 mL of buffer to every 1 × 10⁷ cells for washing, 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 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 effluent (unlabeled cells)	—
	10	Wash the column with buffer three times, 500 μL each time, and add the next wash only after the liquid inside the column has completely drained.	Three times 500 μL of buffer
	11	Combine all effluent fractions into the negative fraction; remove the sorting column from the magnetic field and place it on the collection tube.	—
	12	Add 1 mL of buffer to the column, then 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.

Taking the L Separation Column as an example.

	Steps	Operating procedure	Dosage / Time
Magnetic labeling	1	Count the total number of cells: take the mouse spleen, grind it on a 40 μm sterile filter to prepare a single-cell suspension of the spleen; after red blood cell lysis, proceed with cell counting. 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 buffer. Note:Adjust the volume of buffer proportionally according to the sample volume.	Resuspend cells
	4	Add 10 μL CD11c Nanobeads per 1×10⁷ cells Note:When the cell number increases, proportionally increase the volume of all reagents accordingly	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 with 1 mL of buffer per 1×10⁷ cells, centrifuge at 400×g for 7 minutes, and completely discard the supernatant.	Add 1 mL per 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 flow-through (unlabeled cells).	—
	10	Wash the column with buffer three times, 3 mL each time, and add the next wash only after the liquid in the column has completely drained.	3 × 3 mL of buffer
	11	Combine all eluates into the negative fraction; remove the separation column from the magnetic field and place it over a collection tube.	—
	12	Add 5 mL of buffer to the column, then quickly and firmly push down with the provided 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.

Picture

Validation Data

Using the CD11c Nanobeads Kit, mouse S(L) separation columns, and a cell sorter, CD11c-positive cells were isolated from a single-cell suspension of mouse spleen. Subsequently, fluorescent staining was performed with APC Armenian Hamster Anti-Mouse CD11c Antibody (N418) (STD Cat: S0B5085) and FITC Rat Anti-Mouse I-A/I-E Antibody (M5/114.15.2) (STD Cat: S0B1661), and the live cell population was gated for analysis.

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CD11c Nanobeads Kit, mouse (RUO)