CD45 Nanobeads, human（RUO）

Catalog Number: S0K0001 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 CD45 monoclonal antibody
Capacity	2 mL magnetic beads: suitable for a total cell count of 1×10⁹, and can perform up to 100 separations 1 mL magnetic beads: suitable for 5×10⁸total cells, and can perform up to 50 separations
Principle of Separation	CD45⁺ cells are magnetically labeled using CD45 Nanobeads for human samples. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD45⁺ cells remain retained in the column, while the unlabeled cells flow through; this fraction constitutes the negative fraction after removal of CD45⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD45⁺ cells can be eluted, yielding the positive sorted fraction. To further enhance purity, the positive fraction containing CD45⁺ cells must be subjected to a second separation column.
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

CD45 Nanobeads are designed to positively select or deplete leukocytes from peripheral blood, red blood cell or platelet preparations, lymphoid tissues, tumor tissues, or non-hematopoietic tissues. The CD45 antigen is expressed on all cells of hematopoietic origin, except for red blood cells, platelets, and their precursor cells.

Protocol

Note: This kit does not include sorting 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 according to the sample volume	Resuspend the cells
	4	Add 20 μL of CD45 NanoBeads per 1×10⁷ cells Note:When the cell quantity 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℃ 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 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 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 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 CD45 NanoBeads per 1×10⁷ cells. Note:When the number of cells increases, increase the volumes of all reagents 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 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, 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

CD45+ T cells were enriched from or depleted of mixed cultures of C57BL/6 mouse splenocytes and Jurkat cells (human T-cell leukemia T lymphocytes). Subsequently, …Fluorescent staining was performed using anti-CD45 (clone: 30-F11) conjugated to Alexa Fluor™ 488, and the live cell population was gated for analysis.

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CD45 Nanobeads, human（RUO）