Anti-F4/80 Nanobeads, mouse（RUO）

Catalog Number: S0K0012 Reactivity: Mouse Conjugation: Brand: Starter

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$535 USD

Size: 1mL 2mL

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Product Details

Product Specification

Format	Dextran-coated iron oxide nanomagnets conjugated with an anti-mouse F4/80 monoclonal antibody, in a PBS buffer system containing BSA and Poloxamer 188.
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	Using Anti-F4/80 Nanobeads, mouse anti‑F4/80⁺Cells are subjected to magnetic labeling. Subsequently, the cell suspension is loaded into a separation column positioned within the sorter's magnetic field. The magnetically labeled F4/80…⁺Cells remain retained within the column, while unlabeled cells elute; this fraction constitutes the F4/80-depleted fraction.⁺The negative fraction following cell sorting. After removing the sorting column from the magnetic field, the F4/80‑positive cells that were magnetically captured…⁺Cells can be eluted to obtain the positive‑selection fraction. If higher purity is required, it contains F4/80.⁺The positively sorted cell fraction must be subjected to a second round of sorting on a second 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

F4/80 is a member of the epidermal growth factor (EGF) transmembrane 7 (TM7) family, consisting of a seven-transmembrane molecule and a large extracellular domain containing multiple EGF-like modules. F4/80 is considered one of the most specific cell surface markers of murine macrophages. Constitutive high expression of F4/80 is observed on the surface of most tissue-resident macrophages, including splenic macrophages, microglia in the brain, Kupffer cells in the liver, and Langerhans cells in the skin. Furthermore, F4/80 expression is also regulated according to the physiological state of the cells.

Protocol

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

Taking SSeparation Columnas an example

	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 90 μL of buffer Note:Adjust the volume of buffer proportionally according to the sample volume	Resuspend the cells
	4	Add 10 μL of Anti-F4/80 Nanobeads per 1×10⁷ cells Note:When the number of cells 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 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.

Using 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 per 1×10⁷ cells.Anti-F4/80 Nanobeads. Note:When the cell quantity increases, increase the volume 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 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 three times with 3 mL of buffer each time, adding the next wash only after the reservoir 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 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.

Picture

Validation Data

Enrich or deplete F4/80+ cells from spleen cells of C57BL/6 mice. Stain F4/80+ cells with FITC Rat anti-mouse F4/80 (S-R537) (SDT Cat: S0B5113), and gate on the live cell population for analysis.

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Anti-F4/80 Nanobeads, mouse（RUO）