Mouse CD3/CD28 T Cell Activation Beads, IC-3208
Product Introduction
T cell activation typically requires both TCR/CD3 signaling and CD28 co-stimulatory signaling. TCR/CD3 signaling is initiated when the T cell receptor (TCR) on the T cell surface binds to antigen-presenting molecules on antigen-presenting cells. CD28 co-stimulatory signaling occurs when CD28 on the T cell surface binds to B7 molecules (such as CD80 and CD86) on antigen-presenting cells, providing a second signal that enhances T cell activation, proliferation, and survival.
This product consists of superparamagnetic beads conjugated with Anti-CD3 and Anti-CD28 antibodies, enabling simultaneous activation of TCR and CD28 co-stimulatory pathways without the need for feeder cells (antigen-presenting cells) or exogenous antigens. T cell expansion can be further supported by adding recombinant mouse IL-2 during culture. After activation or expansion, beads can be removed using a magnetic separator.
This product is suitable for T cell activation from mouse splenocytes or T cell subsets including CD3⁺ T cells, CD4⁺T cells, and CD8⁺T cells.
Product Features
1. Dual-Signal Activation: Anti-CD3 and Anti-CD28 antibodies provide both primary and co-stimulatory signals for T cell activation.
2. Feeder-Free System: No antigen-presenting cells or exogenous antigens required.
3. Easy Bead Removal: Magnetic beads can be removed by magnetic separation after activation or expansion.
4. Broad Cell Compatibility: Suitable for mouse splenocytes and CD3⁺, CD4⁺, and CD8⁺ T cell subsets.
5. Reproducible: Uniform 5 µm bead size and consistent antibody coupling ensure reproducible activation.
Specifications
Size: 0.2 mL / 1 mL
Components: Magnetic beads
Ligands: Anti-CD3 antibody, Anti-CD28 antibody
Bead Concentration: 1 ¡Á 10⁸ beads/mL
Bead Diameter: 5 µm
Storage Buffer: PBS, 0.1% BSA, 0.1% Preservative, pH 7.4
Application: Mouse T cell activation and expansion
Storage and Stability
Storage Conditions: Store at 4¡ãC. Do not freeze.
Shelf Life: The product is stable for 2 years from the date of manufacture when stored at 4¡ãC as directed.
Protocol (For Reference Only)
Important: Beads must not be frozen, dried, or subjected to high-speed centrifugation. Avoid prolonged exposure to magnetic fields as this may cause bead aggregation. Use low-binding pipette tips and tubes to minimize bead loss.
Bead Preparation (96-well plate example):
1. Resuspension: Thoroughly resuspend beads by vortexing for >30 seconds or placing on a rotator for >5 minutes.
2. Washing: Transfer 25 µL of beads to a sterile flow tube. Add 1 mL of wash buffer, vortex for 5 seconds or mix by pipetting. Place on a magnetic separator for 3 minutes and discard the supernatant. Repeat washing once with cell culture medium. Resuspend beads in 1 mL of cell culture medium to obtain a concentration of 2.5 ¡Á 10⁶ beads/mL (sufficient for 10 wells of a 96-well plate).
Note: a. Avoid air bubbles during washing. b. If the initial bead volume exceeds 1 mL, use an equal volume of wash buffer. c. Adjust bead volume based on plate format and cell number.
Mouse T Cell Activation (96-well plate example):
1. Adjust T cell concentration to 1 ¡Á 10⁷ cells/mL. Add 25 µL of T cells and 75 µL of culture medium per well (2.5 ¡Á 10⁵ cells/well, 100 µL total).
2. Add 100 µL of washed beads (2.5 ¡Á 10⁵ beads, bead:cell ratio = 1:1, final well volume 200 µL).
3. Incubate at 37¡ãC, 5% CO₂ for the desired activation period.
4. Harvest activated T cells for downstream experiments.
Mouse T Cell Expansion (96-well plate example):
1. Adjust CD3⁺ T cell concentration to 1 ¡Á 10⁶ - 1.5 ¡Á 10⁶ cells/mL. Add washed beads at a bead:cell ratio of 1:1. Incubate in a 37¡ãC, 5% CO₂ incubator.
2. After 2 days, add recombinant mouse IL-2 to a final concentration of 30 U/mL.
Note: a. Observe cell activation and expansion daily. Monitor cell morphology and size. Cell shrinkage or significantly slowed proliferation may indicate exhaustion. b. Avoid additional handling during the first 2 days. After 2 days, change medium or passage when medium turns yellow or cells become overconfluent. c. Replenish recombinant mouse IL-2 to 30 U/mL after each medium change or passage.
3. Count cells regularly. When cell density exceeds 2.5 ¡Á 10⁶ cells/mL, gently pipette to mix and adjust density to 0.5 ¡Á 10⁶ - 1 ¡Á 10⁶ cells/mL.
Precautions
1. When using commercial T cell culture medium for expansion, growth factor concentrations may be adjusted as needed.
2. Use low-binding pipette tips and centrifuge tubes to minimize bead and antibody loss due to adsorption.
3. Avoid high-speed centrifugation, drying, or freezing of beads. Do not leave beads in a magnetic field for prolonged periods as this may cause aggregation and reduce binding activity.
4. Thoroughly resuspend beads before removing from the storage bottle. Handle gently and avoid air bubbles.
5. This product must be used with a magnetic separator.
6. Handle cells gently to avoid mechanical damage.
7. This product is for scientific research by professionals only. Not for use in clinical diagnosis or treatment. Not for use in food or drugs.
8. Wear laboratory protective clothing and disposable gloves when handling this product.
FAQ (Simplified)
Q1: What is the recommended bead-to-cell ratio?
A1: The recommended bead-to-cell ratio is 1:1. This may be adjusted based on experimental requirements.
Q2: How should the beads be stored?
A2: Store at 4¡ãC. Do not freeze. The beads are stable for 2 years under recommended storage conditions.
Q3: How do I remove the beads after activation?
A3: Place the cell-bead mixture on a magnetic separator. Allow beads to separate for 3 minutes, then carefully transfer the supernatant containing activated T cells to a new tube.
Q4: Can these beads be used for human T cells?
A4: No. This product is designed specifically for mouse T cell activation. For human T cells, a human-specific CD3/CD28 activation bead product should be used.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of bead-to-cell ratio and activation conditions is recommended for specific experimental systems.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and disposable gloves when handling this product.
Ordering Information
Catalog Number: IC-3208
Product Name: Mouse CD3/CD28 T Cell Activation Beads
Size: 0.2 mL / 1 mL
Suggested Retail Price:
0.2 mL: CNY ¥1350.00 / USD $135.00 / EUR €162.00 / JPY ¥24300.00
1 mL: CNY ¥5250.00 / USD $525.00 / EUR €630.00 / JPY ¥94500.00
Note: The specific price is subject to the transaction price in the offline contract. This document is provided for technical reference only and does not constitute any form of sales advertisement or invitation to offer. For product details and purchase inquiries, please contact local sales representatives.
