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Streptavidin Nanobeads（120 nm）

Streptavidin Nanobeads（120 nm）

Catalog Number: S0K3012 Reactivity: Human, Mouse Conjugation: Brand: Starter
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$200 USD
$200 USD
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Product Details

Product Specification


Format

Streptavidin-conjugated iron oxide nanomagnetic beads, in a PBS buffer system containing BSA and ProClin 300.
Principle of Separation

Cells are stained with biotinylated primary antibodies or ligands, then magnetically labeled using Streptavidin Nanobeads (120 nm). The cell suspension is subsequently transferred to a polystyrene tube and placed within the magnetic field of a sorter. Under the influence of the magnetic field, the magnetically labeled cells adhere to the tube wall, while unlabeled cells remain in the supernatant. After collecting the supernatant, the magnetically retained cells can be eluted.
Reactivity Human, Mouse
Separation Method Column-Free
Selection Strategy Positive Selection
Stability & Storage

Store protected from light at 2–8 °C; do not freeze.

Background

Streptavidin Nanobeads (120 nm) are designed to isolate cells based on surface markers labeled with biotinylated antibodies or ligands.

Protocol

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

Taking Mouse T column-free negative selection as an example

Note:To achieve better sorting results and cell yield, the amount of magnetic beads needs to be optimized based on different antibody concentrations.

Steps

Operating instructions

Dosage and time

1

Collect the mouse spleen, grind it on a 40 μm sterile filter to prepare a single-cell suspension; after red blood cell lysis, proceed with cell counting;

Note:It is recommended to count cells after staining with AO/PI; splenic cells can be centrifuged at 400 g for 7 minutes;

Prepare the sample

2

Resuspend 1×10 in 100 μL of MagSep Separation Buffer7cells;

Note:Reagents should be adjusted proportionally according to the sample volume; set aside a pre-sorting sample to determine the initial Mouse T ratio;

1×107cells/100 μL

3

Add 10 μL of Mouse T cell Biotin Antibody Cocktail to the sample and gently mix.

Note:For this step, gentle pipetting 2–3 times can be used to mix thoroughly.

10 μL / 100 μL

4

Incubate the antibodies in the cocktail with the cells at room temperature for 10 minutes.

Incubate at room temperature for 10 minutes.

5

Shake for 5–30 seconds to mix the Streptavidin Nanobeads (120 nm).

Mix for 5–30 seconds.

6

Add 5 μL of Streptavidin Nanobeads (120 nm) to the sample;

5 μL/100 μL

7

Gently mix the magnetic beads and cells, and incubate at room temperature for 5 minutes;

Note:For this step, you can use a pipette to gently blow the mixture 2–3 times.

In this step, the magnetic beads may settle at the bottom. You can gently tap to mix them after 2.5 minutes.

Incubate at room temperature for 5 minutes.

8

Add MagSep Separation Buffer to the sample and immediately place it on the ring-shaped magnetic stand to prevent the magnetic beads from settling excessively.

Adjust the total volume to 2.5 mL (using a 5 mL flow cytometry tube).

9

Place the sample on the ring-shaped magnetic stand so that the magnetic beads adhere to the tube wall.

Allow adsorption for 10 minutes.

10

Tilt the magnetic rack and pour the sample into a new collection tube to collect it.

Note: The sorted cells can be centrifuged at 400g for 7 minutes for subsequent culture and analysis.

Cell sorting successful.

Note:For 0.1–1 × 10^8 cells (0.1–1 mL), it is recommended to use a 5 mL single-well magnetic separator (Starter EasyIso Separator) with a total adsorption volume of 2.5 mL;


Picture

Validation Data

T cells were enriched from mouse splenocytes using Streptavidin Nanobeads (120 nm) and a Mouse T cell Biotin Antibody Cocktail. Cells were stained with CD19-Alexa Fluor® 647 (SDT Cat: S0B5445) and CD3-Alexa Fluor® 488 (SDT Cat: S0B8364), and analysis was performed by gating on live cells (Live+), allowing for the determination of the proportion of T cells in the sorted fractions. Flow cytometric analysis revealed that the purity of T cells after this sorting step was 98.4%.