Streptavidin Nanobeads

Catalog Number: S0K0008 Reactivity: Human, Mouse Conjugation: Brand: Starter

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$183 USD

Size: 0.5ml 1mL

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Product Details

Product Specification

Format	Streptavidin-conjugated dextran-coated iron oxide magnetic nanoparticles
Capacity	1 mL magnetic beads: suitable for a total cell count of 1×10⁹, and can perform up to 100 sorting runs 0.5 mL magnetic beads: suitable for 5×10⁸total cells, and can perform up to 50 sorting runs
Principle of Separation	Stain the cells with biotinylated primary antibodies or ligands. Subsequently, magnetically label the cells using Streptavidin Nanobeads. Then, load the cell suspension onto a separation column placed within the separator's magnetic field. The magnetically labeled cells are retained inside the column, while the unlabeled cells pass through it. After removing the column from the magnetic field, the magnetically retained cells can be eluted as the positively selected cell fraction.
Reactivity	Human, Mouse
Separation Method	Column-Based
Selection Strategy	Positive Selection
Stability & Storage	Store protected from light at 2–8°C; do not freeze.

Background

Streptavidin NanobeadsDesigned to isolate cells based on surface markers labeled with biotinylated antibodies or ligands.

Protocol

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

Taking SSeparation Columnas an example

	Steps	Operating procedures	Dosage / Time
Magnetic bead 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 the cells, and label them with biotinylated antibodies according to the manufacturer's instructions. To achieve optimal cell labeling, it is recommended to test different antibody concentrations. Typically, 0.15 to 5 μg of antibody can be added to label 10⁷ mononuclear cells. Incubation at 2–8 °C for 15 minutes is usually sufficient. Note:Ensure thorough mixing of the antibody with the cells by gently pipetting up and down or lightly tapping the bottom of the tube	Biotinylated antibody labeling
	4	Add 1–2 mL of labeling buffer to every 10⁷ cells to wash and remove unbound biotinylated antibodies, then centrifuge at 400×g for 7 minutes. Carefully aspirate the supernatant. Note:The washing steps can be repeated.	Wash the cells.
	5	Resuspend the cell pellet from every 10⁷ total cells in 90 μL of labeling buffer.	Resuspend the cells.
	6	Add 10 μL per 1×10⁷ cells.Streptavidin Nanobeads. Note:When the cell quantity increases, proportionally increase the volume of all reagents accordingly.	Add 10 μL per 1×10⁷ cells.
	7	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes.	Incubate for 15 minutes.
	8	Add 1 mL of buffer per 1×10⁷ cells to wash, centrifuge at 400×g for 7 minutes, and completely aspirate the supernatant.	Add 1 mL per 1×10⁷ cells.
	9	500 μL of buffer can resuspend up to 1×10⁸cells	Resuspend cells
Magnetic separation	10	Place the S-type separation column in the magnetic field of the corresponding magnetic separator, and rinse the column with 500 μL of buffer	Rinse the separation column
	11	Load the cell suspension onto the separation column, and collect the flow-through (unlabeled cells)	—
	12	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
	13	Combine all the flow-through fractions into the negative fraction; remove the separation column from the magnetic field and place it over the collection tube	—
	14	Add 1 mL of buffer to the column, then quickly and firmly push down with the accompanying plunger to elute the magnetically labeled positive cells Note:If higher purity is required, repeat the separation once using a new column	Collect the target cells

In LSeparation ColumnFor 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 the cells and label them with biotinylated antibodies according to the manufacturer's instructions. To achieve optimal cell labeling, it is recommended to test different antibody concentrations. Typically, 0.15 to 5 μg of antibody can be added to label 10⁷ mononuclear cells. Incubation at 2–8 °C for 15 minutes is usually sufficient. Note:Ensure thorough mixing of the antibody with the cells by gently pipetting up and down or lightly tapping the bottom of the tube	Biotinylated antibody labeling
	4	For every 10⁷ cells, add 1–2 mL of labeling buffer to wash the cells and remove unbound biotinylated antibodies, then centrifuge at 400×g for 7 minutes. Completely discard the supernatant. Note:The washing step can be repeated	Washing the cells
	5	Resuspend the cell pellet in 90 μL of labeling buffer for every 10⁷ total cells	Resuspend the cells
	6	Add 10 μL per 1×10⁷ cellsStreptavidin Nanobeads Note:When the cell quantity increases, proportionally increase the volume of all reagents accordingly	Add 10 μL per 1×10⁷ cells
	7	Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes	Incubate for 15 min
	8	Add 1 mL of buffer to wash every 1×10⁷ cells, centrifuge at 400×g for 7 minutes, and completely discard the supernatant	Add 1 mL per 1×10⁷ cells
	9	Use up to 500 μL of buffer to resuspend 1×10⁸Cells	Resuspend the cells
Magnetic separation	10	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
	11	Load the cell suspension onto the separation column and collect the flow-through (unlabeled cells).	—
	12	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
	13	Combine all flow-through fractions into the negative fraction; remove the separation column from the magnetic field and place it over a collection tube.	—
	14	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

Mouse splenocytes were isolated using biotinylated anti-mouse CD3 antibody. Subsequently, the cells were stained with anti-CD3 (clone: 17A2) Alexa Fluor®488, and the live cell population was gated for analysis.

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Streptavidin Nanobeads