Use the CD19 Nanobeads Kit, mouse S or L Separation Column, and a magnetic sorter to enrich or deplete CD19+ cells from mouse spleens. Subsequently, perform fluorescent staining with Alexa Fluor® 647 anti-mouse CD19 Antibody (6D5) and Brilliant Violet 421™ anti-mouse CD3 Antibody (Clone: 17A2), and gate on the live cell population for analysis.
CD19 Nanobeads, mouse（RUO）
CD19 Nanobeads, mouse（RUO）
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$210 USD
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Product Details
Product Details
Product Specification
| Format | Dextran-coated iron tetroxide magnetic nanoparticles conjugated with an anti-mouse CD19 monoclonal antibody |
| Capacity |
1 mL magnetic beads: suitable for a total cell count of 5×108and can perform up to 50 sorting runs. 2 mL magnetic beads: suitable for a total cell count of 1×109and can perform up to 100 sorting runs. |
| Principle of Separation | CD19 Nanobeads, mouse are used to magnetically label CD19⁺ cells. The cell suspension is then loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD19⁺ cells remain retained in the column, while the unlabeled cells elute; this fraction constitutes the negative fraction after removal of CD19⁺ cells. After removing the separation column from the magnetic field, the magnetically captured CD19⁺ cells can be eluted, yielding the positive sorted fraction. |
| Applications in cell sorting | Isolate or deplete CD19+ cells from a single-cell suspension of mouse spleen. |
| Reactivity | 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
Mouse CD19 is encoded by the Cd19 gene and is a type I transmembrane glycoprotein belonging to the immunoglobulin superfamily, serving as a classic marker of the B-cell lineage. This molecule begins to be expressed at the pro-B-cell stage in the bone marrow and remains on the surface of the vast majority of mature B cells, while its expression is downregulated in terminally differentiated plasma 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 method |
Dosage / Time |
|
Magnetic labeling |
1 |
Take the mouse spleen and grind it on a 40 μm sterile filter to prepare a single-cell suspension of splenic cells; after red blood cell lysis, proceed with cell counting. Note:It is recommended to count cells after staining with AO/PI. |
— |
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:The volume of the buffer should be adjusted proportionally according to the sample volume. |
Resuspend the cells. |
|
4 |
Add 20 μL per 1×10⁷ cells.CD19 Nanobeads, mouse. Note:When the cell quantity increases, all reagent volumes should be increased proportionally. |
Add 20 μL to 1×10⁷ cells |
|
5 |
Mix thoroughly, and incubate in the dark at 2–8℃ for 15 minutes |
Incubate for 15 min |
|
6 |
Add 1 mL of buffer to every 1×10⁷ cells for washing, 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 effluent (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. |
Three times 500 μL of buffer |
|
11 |
Combine all effluent fractions into the negative fraction; remove the sorting column from the magnetic field and place it on the collection tube. |
— |
|
12 |
Add 1 mL of buffer to the column, then quickly and firmly push with 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. |
Using LSeparation Columnas an example
Steps |
Operating method |
Dosage / Time |
|
Magnetic labeling |
1 |
Take the mouse spleen, grind it on a 40 μm sterile filter to prepare a single-cell suspension of the spleen; after red blood cell lysis, proceed with cell counting. Note:It is recommended to count cells after staining with AO/PI. |
— |
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 amount of buffer proportionally according to the sample volume. |
Resuspend the cells. |
|
4 |
Add 20 μL per 1×10⁷ cellsCD19 Nanobeads, mouse Note:When the cell number increases, proportionally increase the volume of all reagents accordingly |
Add 20 μL to 1×10⁷ cells |
|
5 |
Mix thoroughly and incubate in the dark at 2–8℃ for 15 minutes |
Incubate for 15 min |
|
6 |
Wash with 1 mL of buffer per 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 in 500 μL of buffer8Cells |
Resuspend the 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 flow-through (unlabeled cells) |
— |
|
10 |
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 |
|
11 |
Combine all eluates into the negative fraction; remove the separation column from the magnetic field and place it over a collection tube. |
— |
|
12 |
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
Picture
Validation Data
DLS
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