CD14 Nanobeads, human（RUO）

Catalog Number: S0K0004 Reactivity: Human Conjugation: Brand: Starter

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$283 USD

Size: 1mL 2mL

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Product Details

Product Specification

Format	Dextran-coated iron tetroxide nanomagnetic beads coupled with an anti-human CD14 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	CD14⁺ cells are magnetically labeled using CD14 Nanobeads. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD14⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD14⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD14⁺ cells can be eluted, yielding the positive sorted fraction.
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

CD14 Nanobeads are used for the positive selection or depletion of human monocytes and macrophages from umbilical cord blood or peripheral blood mononuclear cells (PBMCs), as well as from pleural effusions, ascitic fluid, synovial fluid, or various tissues such as the spleen and lymph nodes. The CD14 antigen is part of the LPS receptor complex. Because CD14 lacks a cytoplasmic domain, antibody binding to CD14 does not trigger signal transduction. CD14 is strongly expressed on most monocytes and macrophages and weakly expressed on neutrophils and some myeloid dendritic cells.

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 80 μL of buffer to resuspend every 1×10⁷ cells Note:Adjust the amount of buffer proportionally based on the sample volume	Resuspend the cells
	4	Add 20 μL of CD14 NanoBeads per 1×10⁷ cells Note:When the cell quantity increases, all reagent volumes should be increased proportionally	Add 20 μL per 1×10⁷ cells
	5	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes	Incubate for 15 minutes
	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 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 with buffer three times, 500 μL each time, and add 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 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 CD14 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 and incubate in the dark at 2–8°C for 15 minutes.	Incubate for 15 minutes.
	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, each time using 3 mL; 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 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 CD14+ cells from human PBMCs (human peripheral blood mononuclear cells). Subsequently, perform fluorescent staining with anti-CD14 (clone: 61D3) APC and gate on the live cell population for analysis.

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CD14 Nanobeads, human（RUO）