AcnoviaBeads Human CD3 (Cat. No. AC68965) is composed of 4.5 μm magnetic beads conjugated with anti-human CD3 antibodies, which can easily separate or remove human CD3⁺ T cells from peripheral blood, bronchial lavage, cell culture, or various tissues such as lymphoid, nasal, and tumor tissue. T cells isolated by AcnoviaBeads Human CD3 have been used for various studies, such as T cell cytotoxicity, T cell activation, signal transduction and so on.

Catalog：AC68965

Reactivity：Human

Concentration：2×10⁸ beads/mL

Particle sise：4.5 μm

Endotoxin：<1 EU/mg

Usage：Separate or remove human CD3⁺T cells from peripheral blood, bronchial lavage, cell culture, or various tissues such as lymphoid, nasal, and tumor tissue

Formulation：phosphate buffered saline (PBS), containing Bovine Serum Albumin (BSA), pH 7.4

Stability：24 months

Storage：2℃ to 8℃,Do not freeze

Wash AcnoviaBeads Human CD3

1. Resuspend the AcnoviaBeads Human CD3 (beads) in the tube (vortex for >30 sec, or tilt and rotate for 5min).

2. Transfer the beads with a desired volume into a tube.

3. Add equal volume of PBS (0.1%BSA), if the beads volume is less than 1m L, add 1mL PBS (0.1%BSA) for resuspension.

4. Place the tube in a magnet for 1 min, and then discard the supernatant.

5. Remove the tube from the magnet, then use the same volume of PBS (0.1%BSA) to resuspend the beads.

Separate CD3⁺ T cells

This protocol is based on 1 mL (1x10⁷) PBMCs. If the volume is less than 1m L, add PBS (0.1%BSA) for resuspension. If the volume is larger than 1 mL, scale up all volumes in corresponding proportions.

1. Transfer 1mL (1x10⁷) PBMCs and add 50 uL washed beads.

2. Rotate the samples with a speed of 50~120 rpm/min, and incubate them at room temperature for 30 min.

3. Dilute the mixture of beads and cells with cell culture media or PBS (0.1%BSA) to ensure the separation volume for magnetic selection. Following this, place the tube on the magnet for 1~2min.

4. Remove the supernatant, then resuspend the mixture of beads and cells with PBS (0.1%BSA), and place the tube on the magnet for 1~2min, While the tube is still in the magnet, carefully remove and discard the supernatant.

5. Wash the bead-bound CD3⁺ T cells for 2-3 times, and then resuspend the mixture of beads and cells with preferred cell culture media. Keep the cells on 2°C to 8°C until further use in downstream applications.

Materials Required

1. Magnet

2. Mixer allowing tilting and rotation of tubes

Notices and Tips

1. It is forbidden to place the product in a magnetic field for a long time to avoid magnetic bead agglomeration, resulting in reduced binding activity.

2. When this product is stored unopened at 2-8 ℃, it is stable until the expiration date indicated on the label. Store the opened vials at 2-8 ℃ to avoid bacterial contamination.

3. Keep the product in liquid suspension during storage and all handling steps, as drying can cause performance degradation, resuspend before use.

4. This product is for research purposes only.