Human CD3⁺ Cell Removal Beads (C008B-268531)

Human CD3⁺ Cell Removal Beads are generated by covalently coupling anti‑human CD3 antibody to magnetic beads. They can be used for rapid and efficient removal of CD3⁺ cells from human peripheral blood mononuclear cells (PBMCs) or from red blood cell‑lysed human peripheral blood white blood cells (WBCs).ApplicationsRemoval of CD3⁺ cells from human PBMCs or WBCs.Beads Volume (suspension)Total Cell Number250 μL5 × 10⁸ cells1 mL2 × 10⁹ cellsProtocolWhen the sample is PBMC or WBC:Prepare single‑cell suspensionCount the obtained human PBMCs or human WBCs. Resuspend 1×10⁸ cells in 450 μL of Isolation Buffer.Note: Isolation buffer is PBS containing 2 mM EDTA and 2% fetal bovine serum (FBS). FBS may be replaced with 0.5% BSA or HSA. The buffer must be pre‑filtered through a 0.22 μm filter.Incubate cells with beadsAdd the 450 μL cell suspension prepared in step 1 to the bottom of a 1.5 mL centrifuge tube. Then add 50 μL of washed and resuspended Human CD3⁺ Cell Removal Beads (the beads must be washed before use: select a pipette with an appropriate volume range, thoroughly resuspend the beads by pipetting, avoiding bubbles. Transfer the required volume of beads to a 1.5 mL centrifuge tube, add 1 mL of Isolation Buffer, centrifuge at 10,000 g for 1 min or use a magnetic separator for 3 min, then discard the supernatant. Add 1 mL of Isolation Buffer, wash the beads once more, and then resuspend the beads in the same volume of Isolation Buffer as originally taken. For example, if 50 μL of beads are taken, resuspend them in 50 μL of Isolation Buffer after washing). Mix the liquid in the tube by pipetting, and incubate at room temperature for 30 min using a rotator.Note: For other cell numbers, adjust the cell volume and the amount of Human CD3⁺ Cell Removal Beads proportionally. For fewer than 5×10⁷ cells, adjust the cell suspension volume to 225 μL and add 25 μL of Human CD3⁺ Cell Removal Beads.Magnetic separationAfter incubation, add Isolation Buffer to bring the total volume to 3 mL (the liquid can be transferred from the 1.5 mL centrifuge tube to a FACS tube in several steps). Mix thoroughly by pipetting up and down 10 times (avoid vigorous shaking or inverting). Then place the FACS tube on a magnetic separator and let stand for 5 min.Collect target cellsAfter magnetic separation, the target cells are in the supernatant. Carefully collect the supernatant (avoid taking up any beads), centrifuge at 500 g for 5 min, discard the supernatant, and collect the cells.Resuspend for further useWash the cells if required for your experiment, or centrifuge and resuspend them in the desired buffer or culture medium for subsequent molecular or cell biology experiments.Removal PerformanceCD3⁺ cells were removed from human WBCs and human PBMCs. Cells before and after removal were stained with APC anti‑human CD3 antibody (clone OKT3) and PE anti‑human CD4 antibody (clone RPA‑T4) and analyzed by flow cytometry. The results are as follows:WBCs: CD3⁺ cell purity before and after removal was 16.39% (mean±SD: 19.34%±4.16%) and 0.1% (mean±SD: 0.075%±0.04%), respectively.PBMCs: CD3⁺ cell purity before and after removal was 53% (mean±SD: 47.51%±9.67%) and 0.28% (mean±SD: 0.42%±0.25%), respectively.PrecautionsAvoid freezing the beads during use and storage.Low‑retention pipette tips and centrifuge tubes are recommended to avoid loss of beads due to adsorption.Before taking the beads, select a pipette with an appropriate volume range, thoroughly resuspend the beads by pipetting, and avoid generating bubbles during pipetting.For research use only.

Specifications & Purity: Human CD3⁺ Cell Removal Beads are generated by covalently coupling anti‑human CD3 antibody to magnetic beads. They can be used for rapid and efficient removal of CD3⁺ cells from human peripheral blood mononuclear cells (PBMCs) or from red blood cell‑lysed human peripheral blood white blood cells (WBCs).ApplicationsRemoval of CD3⁺ cells from human PBMCs or WBCs.Beads Volume (suspension)Total Cell Number250 μL5 × 10⁸ cells1 mL2 × 10⁹ cellsProtocolWhen the sample is PBMC or WBC:Prepare single‑cell suspensionCount the obtained human PBMCs or human WBCs. Resuspend 1×10⁸ cells in 450 μL of Isolation Buffer.Note: Isolation buffer is PBS containing 2 mM EDTA and 2% fetal bovine serum (FBS). FBS may be replaced with 0.5% BSA or HSA. The buffer must be pre‑filtered through a 0.22 μm filter.Incubate cells with beadsAdd the 450 μL cell suspension prepared in step 1 to the bottom of a 1.5 mL centrifuge tube. Then add 50 μL of washed and resuspended Human CD3⁺ Cell Removal Beads (the beads must be washed before use: select a pipette with an appropriate volume range, thoroughly resuspend the beads by pipetting, avoiding bubbles. Transfer the required volume of beads to a 1.5 mL centrifuge tube, add 1 mL of Isolation Buffer, centrifuge at 10,000 g for 1 min or use a magnetic separator for 3 min, then discard the supernatant. Add 1 mL of Isolation Buffer, wash the beads once more, and then resuspend the beads in the same volume of Isolation Buffer as originally taken. For example, if 50 μL of beads are taken, resuspend them in 50 μL of Isolation Buffer after washing). Mix the liquid in the tube by pipetting, and incubate at room temperature for 30 min using a rotator.Note: For other cell numbers, adjust the cell volume and the amount of Human CD3⁺ Cell Removal Beads proportionally. For fewer than 5×10⁷ cells, adjust the cell suspension volume to 225 μL and add 25 μL of Human CD3⁺ Cell Removal Beads.Magnetic separationAfter incubation, add Isolation Buffer to bring the total volume to 3 mL (the liquid can be transferred from the 1.5 mL centrifuge tube to a FACS tube in several steps). Mix thoroughly by pipetting up and down 10 times (avoid vigorous shaking or inverting). Then place the FACS tube on a magnetic separator and let stand for 5 min.Collect target cellsAfter magnetic separation, the target cells are in the supernatant. Carefully collect the supernatant (avoid taking up any beads), centrifuge at 500 g for 5 min, discard the supernatant, and collect the cells.Resuspend for further useWash the cells if required for your experiment, or centrifuge and resuspend them in the desired buffer or culture medium for subsequent molecular or cell biology experiments.Removal PerformanceCD3⁺ cells were removed from human WBCs and human PBMCs. Cells before and after removal were stained with APC anti‑human CD3 antibody (clone OKT3) and PE anti‑human CD4 antibody (clone RPA‑T4) and analyzed by flow cytometry. The results are as follows:WBCs: CD3⁺ cell purity before and after removal was 16.39% (mean±SD: 19.34%±4.16%) and 0.1% (mean±SD: 0.075%±0.04%), respectively.PBMCs: CD3⁺ cell purity before and after removal was 53% (mean±SD: 47.51%±9.67%) and 0.28% (mean±SD: 0.42%±0.25%), respectively.PrecautionsAvoid freezing the beads during use and storage.Low‑retention pipette tips and centrifuge tubes are recommended to avoid loss of beads due to adsorption.Before taking the beads, select a pipette with an appropriate volume range, thoroughly resuspend the beads by pipetting, and avoid generating bubbles during pipetting.For research use only.

Grade: BioReagent

Storage Temperature: Store at 2-8°C;Do not freeze

Shipped in: Wet ice;Do not freeze

UPC:

12352206

Weight:

1.09 Ounces

HazmatClass:

No

Quantity:

1

Unit of Measurement:

EA

WeightUOM:

LB

MPN:

R1522426-1ml

Country of Origin:

China

Product Size:

1ml

Harmonized Tariff Code:

2931.90.90