CD11b+ cells are enriched or depleted from human and mouse peripheral blood leukocytes using CD11b NanoBeads, S or L Separation Columns, and a magnetic separator. Subsequently, fluorescent staining is performed with anti-CD11b (clone: M1/70) Brilliant Violet 421™, and the live cell population is gated for analysis.
CD11b NanoBeads, human and mouse（RUO）
CD11b NanoBeads, human and mouse（RUO）
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Product Details
Product Details
Product Specification
| Format | Dextran-coated iron oxide nanomagnetic beads conjugated with anti-human and anti-mouse CD11b monoclonal antibodies, in a PBS buffer system containing BSA and Poloxamer 188. |
| Capacity | 1. For human whole blood or mouse bone marrow cells: Add 20 μL of nanobeads per 10⁷ cells, allowing for up to 100 separations. 2. For human PBMCs: Add 6 μL of nanobeads per 10⁷ cells, allowing for up to 333 separations. 3. For mouse splenocytes: Add 2 μL of nanobeads per 10⁷ cells, allowing for up to 1,000 separations. |
| Principle of Separation | AdoptCD11b NanoBeads, human and mouseMagnetic labeling of CD11b⁺ cells. Subsequently, the cell suspension is loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD11b⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD11b⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD11b⁺ cells can be eluted, yielding the positive sorted fraction. |
| Reactivity | Human, Mouse |
| 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
CD11b NanoBeads, human and mouse are designed to isolate human and murine cells based on the expression of the CD11b antigen. In humans, CD11b is highly expressed on myeloid cells and weakly expressed on NK cells and some activated lymphocytes. In mice, the CD11b antigen is expressed on monocytes/macrophages, with lower levels observed on granulocytes, NK cells, CD5+ B1 cells, and certain subsets of dendritic cells. This CD11b antibody (Mac-1 α chain; integrin αM chain) binds to the 170 kDa αM subunit within the CD11b/CD18 heterodimer (Mac-1, αMβ2 integrin). It functions as a receptor for complement components (C3bi), fibrinogen, or coagulation factor X.
Protocol
Note: This kit does not include sorting buffer; please prepare it yourself. The corresponding Stater product number isS0D3018
Taking S as an exampleSeparation ColumnFor instance
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 |
When processing human whole blood or mouse cells: resuspend the cell pellet with 80 μL of buffer per 1×10⁷ total cells; When processing PBMCs: resuspend the cell pellet with 94 μL of buffer per 1×10⁷ total cells Note:The amount of buffer should be adjusted proportionally according to the sample volume |
Resuspend the cells |
|
4 |
When processing human whole blood or mouse cells: add 20 μL of CD11b NanoBeads per 1×10⁷ total cells; When handling PBMCs: add 6 μL of CD11b NanoBeads per 1×10⁷ total cells. Note:When the cell number increases, proportionally increase the volume of all reagents accordingly. |
Add magnetic beads |
|
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 discard the supernatant. |
Add 1 mL to 1×10⁷ cells |
|
7 |
Resuspend up to 1×10 with 500 μL of buffer8Cells |
Resuspend the 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 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 1 mL of buffer to the column, then quickly and firmly push down with the provided 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. |
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 |
When processing human whole blood or mouse cells: resuspend the cell pellet with 80 μL of buffer per 1×10⁷ total cells; When processing PBMCs: Resuspend the cell pellet in 94 μL of buffer per 1×10⁷ total cells. Note:Adjust the volume of the buffer proportionally according to the sample volume. |
Resuspend the cells |
|
4 |
When processing human whole blood or mouse cells: Add 20 μL of CD11b NanoBeads per 1×10⁷ total cells; When processing PBMCs: Add 6 μL of CD11b NanoBeads per 1×10⁷ total cells. Note:When the number of cells increases, increase the volumes of all reagents proportionally. |
Add magnetic beads |
|
5 |
Mix thoroughly and incubate at 2–8℃ in the dark for 15 minutes. |
Incubate for 15 minutes |
|
6 |
For every 1×10⁷ cells, add 1 mL of buffer for washing, centrifuge at 400×g for 7 minutes, and completely discard the supernatant. |
Add 1 mL per 1×10⁷ cells |
|
7 |
Resuspend up to 1×10 with 500 μL of buffer8Cells |
Resuspend cells |
|
Magnetic sorting |
8 |
Place the L-shaped sorting column in the magnetic field of the corresponding magnetic separator, and rinse the column with 3 mL of buffer |
Rinse the sorting column |
9 |
Load the cell suspension onto the sorting column, and collect the flow-through (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 the flow-through fractions 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, then quickly and firmly push down with the accompanying 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
Picture
Validation Data
