Human CD66b+ Cell Positive Selection Kit
MCE Human CD66b+ Cell Positive Selection Kit is designed for the efficient isolation of CD66b+ cells from human peripheral blood single-cell suspensions. CD66b+ cells are first specifically labeled with the CD66b Capture Antibody and subsequently captured using Releasable Magnetic Beads. Following magnetic separation, the beads are detached from the cell surface using Magnetic Beads Release Buffer, resulting in highly purified CD66b+ cells free of residual magnetic bead labeling.
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Storage :
4°C, 2 years
Do not freeze the magnetic beads
- Manual (3385KB)
- COA
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
MCE Human CD66b+ Cell Positive Selection Kit is designed for the efficient isolation of CD66b+ cells from human peripheral blood single-cell suspensions. CD66b+ cells are first specifically labeled with the CD66b Capture Antibody and subsequently captured using Releasable Magnetic Beads. Following magnetic separation, the beads are detached from the cell surface using Magnetic Beads Release Buffer, resulting in highly purified CD66b+ cells free of residual magnetic bead labeling.
Features of MCE Human CD66b+ Cell Positive Selection Kit:
1. Simple and Rapid: No separation columns are required. Cell isolation can be completed using only a magnetic separator.
2. High Purity: Optimized magnetic capture enables efficient enrichment, yielding up to 98% purity of isolated CD66b+ cells.
3. High Cell Viability: The isolated CD66b+ cells are free of magnetic bead labeling, maintaining high viability and normal cellular functions for downstream applications.
Protocol
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: Isolation of human CD66b+ cells from a human peripheral-blood single-cell suspension
1. Prepare a single-cell suspension from a human peripheral-blood single-cell suspension. Remove red blood cells as appropriate, resuspend the cells in PBS, 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. Transfer 100 μL of cell suspension (1 × 107 cells) to the bottom of a sterile centrifuge tube. Add 2 μL CD66b Capture Antibody, mix thoroughly, and incubate at 4°C for 10 min.
Note: Scale the amount of capture antibody proportionally for larger cell numbers. For fewer than 1 × 107 cells, adjust the cell suspension volume to 100 μL.
4. Preparation of Releasable Magnetic Beads: thoroughly resuspend the beads and transfer 20 μL 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.
5. After antibody incubation, add 20 μL of pre-washed Releasable Magnetic Beads, mix thoroughly, and incubate at 4°C for 10 min.
6. Add 2.5 mL Isolation Buffer and mix gently. Place the tube on a magnetic separation rack for 5 min.
7. While the tube remains on the magnetic rack, carefully discard the supernatant. Remove the tube from the rack, add 2.5 mL Isolation Buffer, and thoroughly resuspend the bead-cell complexes.
8. Place the tube on the magnetic rack again for 5 min and discard the supernatant. Repeat Steps 7-8 for a total of two washes.
9. Remove the tube from the magnetic rack, add 400 μL Magnetic Beads Release Buffer, gently resuspend the bead-cell complexes, and incubate at room temperature for 10 min. Mix gently every 2-3 min during incubation.
10. Add 2.5 mL Isolation Buffer, mix gently, and place the tube on the magnetic rack for 5 min.
11. While keeping the tube on the magnetic rack, carefully collect the supernatant into a new sterile centrifuge tube. The supernatant contains bead-free human CD66b+ cells. Centrifuge at 300 × g for 5 min, discard the supernatant, and collect the cells.
12. Wash the cells as required and resuspend them in an appropriate buffer or culture medium. The isolated cells may be used 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 high-speed centrifugation, 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. /p>
Components
| Components | HY-K0358-0.2 mL (For 108 cells) | HY-K0358-1 mL (For 109 cells) |
|---|---|---|
| CD66b Capture Antibody | 20 μL | 200 μL |
| Releasable Magnetic Beads | 0.2 mL | 1 mL × 2 |
| Magnetic Beads Release Buffer | 4 mL | 40 mL |
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