CD66b Nanobeads, human（RUO）

Catalog Number: S0K0023 Reactivity: Human Conjugation: Brand: Starter

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$450 USD

Size: 1 mL 2 mL

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Product Details

Product Specification

Format	Dextran-coated iron tetroxide nanomagnetic beads coupled with an anti-human CD66b monoclonal antibody
Capacity	1 mL magnetic beads: suitable for a total cell count of 5 × 10⁸and can perform up to 50 sorting runs. 2 mL magnetic beads: suitable for a total cell count of 1 × 10⁹and can perform up to 100 sorting runs.
Principle of Separation	CD66b Nanobeads are used to magnetically label human CD66b⁺ cells. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD66b⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD66b⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD66b⁺ cells can be eluted, yielding the positive sorted fraction.
Applications in cell sorting	Isolate or deplete CD66b+ cells from human whole blood (following red blood cell lysis).
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

CD66b is a glycosylphosphatidylinositol (GPI)-anchored protein with a molecular weight of 95–100 kDa, also known as CD67, CGM6, and NCA-95. This molecule is expressed on the surface of neutrophils and eosinophils but is not expressed on basophils or lymphocytes.

Protocol

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

Taking S as an exampleSeparation Columnas an example

	Steps	Operating method	Dosage / Time
Magnetic labeling	1	Count the total number of cells after lysing red blood cells in human whole blood 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 80 μL of buffer Note:The amount of buffer should be adjusted proportionally according to the sample volume.	Resuspend the cells
	4	Add 20 μL per 1×10⁷ cellsCD66b Nanobeads, human Note:When the cell quantity increases, all reagent volumes should be increased proportionally; if you need to improve cell yield, you can appropriately increase the amount of magnetic beads.	Add 20 μL per 1×10⁷ cells
	5	Mix thoroughly, incubate at 2–8°C 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	1×10⁷ cells plus 1 mL
	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, and add the next wash only after the liquid inside 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, then quickly and firmly press 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 after lysing red blood cells in human whole blood 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 80 μL of buffer. Note:Adjust the amount of buffer proportionally according to the sample volume.	Resuspend the cells.
	4	Add 20 μL per 1 × 10⁷ cells.CD66b Nanobeads, human Note:When the cell number increases, all reagent volumes should be increased proportionally; to improve cell yield, the amount of magnetic beads can be appropriately increased.	Add 20 μL to 1×10⁷ cells
	5	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes	Incubate for 15 min
	6	Add 1 mL of buffer per 1×10⁷ cells to wash, 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 in the column has completely drained.	3 × 3 mL of buffer
	11	Combine all eluates as 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:If higher purity is required, use a new column and repeat the separation once.	Collect the target cells.

Picture

Validation Data

Use CD66b Nanobeads, a human S or L Separation Column, and a magnetic separator to enrich or deplete CD66b+ cells from human whole blood. Subsequently, perform fluorescent staining with Alexa Fluor® 488 Mouse Anti-Human CD66b Antibody (G10F5) (STD Cat: S0B8109) and APC anti-human CD14 Antibody (Clone: HCD14), and gate on the live cell population for analysis.

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CD66b Nanobeads, human（RUO）