Human CD3 Cell Depletion Magnetic Beads (Positive Selection)
MCE Human CD3 Cell Depletion Magnetic Beads (Positive Selection) are magnetic beads covalently coupled with anti-human CD3 antibodies, enabling the specific binding and magnetic removal of CD3+ cells from single-cell suspensions derived from human peripheral blood mononuclear cells (PBMCs) or human peripheral blood leukocytes (WBCs). This allows efficient enrichment of the non-CD3+ cell population.
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Storage :
4°C, 2 years. Do not freeze the magnetic beads.
- Manual (2717KB)
- COA
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
MCE Human CD3 Cell Depletion Magnetic Beads (Positive Selection) are magnetic beads covalently coupled with anti-human CD3 antibodies, enabling the specific binding and magnetic removal of CD3+ cells from single-cell suspensions derived from human peripheral blood mononuclear cells (PBMCs) or human peripheral blood leukocytes (WBCs). This allows efficient enrichment of the non-CD3+ cell population.
This product is suitable for single-cell suspensions prepared from human PBMCs or WBCs following red blood cell lysis.
Features of MCE Human CD3 Cell Depletion Magnetic Beads (Positive Selection):
1. Simple and Rapid: CD3+ cell enrichment or depletion can be completed within approximately 30 min.
2. High Purity: Achieves up to 99% depletion of CD3+ cells.
3. High Cell Viability: The isolated non-CD3+ cells are free of antibody and magnetic bead labeling, maintaining high viability and normal cellular functions for downstream applications.
Protocols
Recommended Buffer (Not Provided)
| Buffer | Composition |
|---|---|
| Isolation Buffer | PBS, 0.5% BSA, 2 mM EDTA, pH 7.0-7.4 |
Note: a. BSA may be replaced with human serum albumin (HSA) or 2% fetal bovine serum (FBS);
b. It is recommended to prepare the Isolation Buffer with water for injection, sterilize it through a 0.22 μm filter, and store it at 4°C.
Example: Depletion of CD3+ cells from a human peripheral blood mononuclear cells (PBMCs) or other human cells single-cell suspension
1. Prepare a human peripheral blood mononuclear cells (PBMCs) or other human cells single-cell suspension and perform red blood cell lysis as appropriate for the sample type. Filter through a 70 μm cell strainer and count the cells. Centrifuge at 500 × g for 5 min and discard the supernatant.
2. Resuspend the cells in Isolation Buffer and adjust the cell density to 1 × 108 cells/mL.
3. Thoroughly resuspend the CD3 Cell Depletion Magnetic Beads. Transfer 20 μL beads to a 1.5 mL microcentrifuge tube, add 1 mL Isolation Buffer, mix, centrifuge at 10,000 × g for 1 min, and discard the supernatant. Repeat the wash 1-2 times and finally resuspend the beads in 20 μL Isolation Buffer.
4. Transfer 100 μL cell suspension (1 × 107 cells) to the bottom of a sterile centrifuge tube. Add 20 μL of pre-washed beads, mix thoroughly, and incubate at 4°C for 15 min.
Note: Scale the bead volume proportionally for larger cell numbers. For fewer than 1 × 107 cells, adjust the suspension volume to 100 μL and add 20 μL beads.
5. After incubation, add 2.5 mL Isolation Buffer and mix gently.
6. Place the tube on a magnetic separation rack and allow it to stand for 5 min.
7. While keeping the tube on the magnetic rack, carefully transfer the supernatant to a new sterile centrifuge tube. The supernatant contains the CD3+-depleted cell population.
8. To increase depletion efficiency, the collected supernatant may be subjected to a second round of bead treatment by repeating Steps 3-7.
9. Centrifuge at 300 × g for 5 min, discard the supernatant, and collect the cells. Wash and resuspend the cells as required for downstream experiments.
Storage
4°C, 2 years.
Do not freeze the magnetic beads.
Attention
1. Use low-retention pipette tips and tubes whenever possible to minimize bead and antibody loss due to surface adsorption.
2. Avoid drying or freezing of the magnetic beads during storage and handling.
3. Do not leave the magnetic beads in a magnetic field for extended periods, as this may cause bead aggregation and reduced binding activity.
4. Thoroughly resuspend the beads before use. Handle gently throughout the procedure and avoid generating excessive bubbles.
5. This product must be used with a compatible magnetic separator.
6. Perform all procedures gently to minimize mechanical stress and maintain cell viability.
7. This product is for R&D use only, not for drug, household, or other uses.
8. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K0356-0.25 mL (For 5 × 108 cells) | HY-K0356-1 mL (For 2 × 109 cells) |
|---|---|---|
| Human CD3 Antibody Magnetic Beads | 0.25 mL | 1 mL |
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