Dynabeads™ Mouse T-Activator CD3/CD28 for T-Cell Expansion and Activation
Dynabeads™ Mouse T-Activator CD3/CD28 for T-Cell Expansion and Activation
Gibco™

Dynabeads™ Mouse T-Activator CD3/CD28 for T-Cell Expansion and Activation

Dynabeads Mouse T-Activator CD3/CD28 beads are 4.5-μm superparamagnetic beads covalently coupled with an optimized combination of anti-mouse CD3 and anti-mouseRead more
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Catalog NumberVolume (Metric)
11456D0.4 mL
11452D2 mL
11453D5 x 2 mL
Catalog number 11456D
Price (USD)
202.00
Each
Add to cart
Volume (Metric):
0.4 mL
Price (USD)
202.00
Each
Add to cart

Dynabeads Mouse T-Activator CD3/CD28 beads are 4.5-μm superparamagnetic beads covalently coupled with an optimized combination of anti-mouse CD3 and anti-mouse CD28 antibodies and intended for ex vivo activation and expansion/proliferation of mouse T cells. T cells are one of the most important cells in the immune system and are responsible for fighting off infections and cancer and are therefore intensively used in many cancer and disease studies, including CAR-T cell research.

Benefits of Dynabeads Mouse T-Activator CD3/CD28 beads include:
Easy—activate T cells without the need for feeder cells (antigen-presenting cells)
Efficient—reproducible method and high T-cell expansion rate (up to 1000-fold)
No contamination—using magnetic beads avoids contamination of soluble antibodies or mitogens
Functional­—activated T cells retain the in vivo-like function

Simple and fast procedure
Dynabeads Mouse T-Activator CD3/CD28 beads offer a simple method for the activation and expansion of CD3+ T cells, CD4+ T helper cells or regulatory T cells, and CD8+ T cytotoxic cells isolated from the spleen, lymph nodes, mononuclear cells (MNC’s), or whole blood. Isolated T cells are activated by adding the beads and recombinant mouse or human IL-2 (rIL-2) to the cell culture. Optimal expansion is performed when appropriate T-cell medium is used, such as CTS OpTmizer T Cell Expansion SFM or Advanced RPMI 1640 Medium with added supplements according to the manual.

Activation of mouse T cells
Short-time activation: activation of T cells with Dynabeads Mouse T-Activator CD3/CD28 for a few hours to a few days. This procedure does not expand the cells, and proteomics expression/detection and nucleic acid assays can be performed. The cell:bead complexes cannot be used in flow cytometry in this phase.

Long-term activation/expansion: activation of T cells with Dynabeads Mouse T-Activator CD3/CD28 for three days to a few weeks to obtain an expansion of the T cells. When experiencing reduced expansion or cell shrinking, the T cells can be restimulated with the Dynabeads Mouse T-Activator CD3/CD28 beads. The beads can be removed with a DynaMag magnet and used in applications like cell:cell interaction studies, flow cytometry, proteomics, or nucleic acid-based assays.

Pre-isolate the mouse T cells
To obtain the most optimal activation and expansion, pure T cells can be isolated directly from single cell suspension from mouse spleen or lymph nodes with Dynabeads magnetic beads. For positive isolation of mouse CD3, CD4, or CD8 T cells with a release, use our Dynabeads Mouse FlowComp products. For negative isolation of mouse CD3, CD4, or CD8 T cells, use our Dynabeads Untouched Mouse cell products.

Alternative human activation products, from research to clinic
• For activation of polyclonal T cells, use Dynabeads Human T Activator CD3/CD28
• For activation of regulatory T cells (Tregs), use Dynabeads CD3/CD28 Treg Expander
• For activation of antigen-specific T cells, use Dynabeads Human T Activator CD3/CD28/CD137
• For ex vivo activation and expansion of T cells for cell-based therapy, use the aseptically manufactured CTS Dynabeads CD3/CD28

Commercial supply
Our manufacturing sites are ISO 13485–certified. Bead characteristics and manufacturing conditions help ensure consistency and batch reproducibility making them an ideal choice for commercial supply. If you are utilizing Dynabeads Mouse T-Activator CD3/CD28 beads in a commercial product or service or need a larger bulk volume of this product, please contact us at oemdynal@lifetech.com or read more on our Dynabeads OEM page.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cell TypeMouse T-cells
Certifications/ComplianceISO9001 and ISO13485
ClonalityMonoclonal
Concentration4 x 10 7 Beads/mL
DescriptionMonoclonal antibodies covalently bound to DynaBeads magnetic Beads
ColorBrown
For Use With (Application)Activation and Expansion of T-cells
MaterialMagnetic Polystyrene
Product TypeCell Expansion and Activation Kit
For Use With (Equipment)DynaMag™ Magnet
Host SpeciesHamster
Isolation TechnologyMagnetic Beads
Ligand TypeAntibody
Product LineDynaBeads™
ReactivityMouse
Sample TypeCell Cultures, Lymph Node, Spleen
Shelf Life36 months from date of manufacture
Shipping ConditionAmbient Temperature
Surface FunctionalityEpoxy
Target SpeciesMouse T-cells
UniformityMonosized 4.5 μm (CV <5%)
Volume (Metric)0.4 mL
Diameter (Metric)4.5 μm
Regulatory StatusFor Research Use Only
Unit SizeEach
Contents & Storage
DynaBeads Human T-Activator CD3/CD28 are supplied in PBS, pH 7.4, with 0.1% HSA. Store at 2°C to 8°C.

Frequently asked questions (FAQs)

My Dynabeads magnetic beads are not pelleting well with the magnet. Do you have any suggestions for me?

Please review the following possibilities for why your Dynabeads magnetic beads are not pelleting:

- The solution is too viscous.
- The beads have formed aggregates because of protein-protein interaction.

Try these suggestions: - Increase separation time (leave tub on magnet for 2-5 minutes)
- Add DNase I to the lysate (~0.01 mg/mL)
- Increase the Tween 20 concentration to ~0.05% of the binding and/or washing buffer.
- Add up to 20 mM beta-merecaptoethanol to the binding and/or wash buffers.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

I have a long double-stranded DNA fragment I would like to isolate. What product do you recommend?

For biotin-labeled DNA that is less than 1 kb, we recommend you use Dynabeads M270 Streptavidin (Cat. No. 65305) and MyOne C1 magnetic beads (Cat. No. 65001). We recommend our Dynabeads KilobaseBINDER Kit (Cat. No. 60101), which is designed to immobilize long (>1 kb) double-stranded DNA molecules. The KilobaseBINDER reagent consists of M-280 Streptavidin-coupled Dynabeads magnetic beads along with a patented immobilization activator in the binding solution to bind to long, biotinylated DNA molecules for isolation. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/immobilisation-of-long-biotinylated-dna-fragments.html) for more information in regards to long biotinylated DNA fragment isolation.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

Can I use Dynabeads magnetic beads to isolate single-stranded DNA templates?

Yes, Dynabeads magnetic beads can be used to isolate single-stranded DNA. Streptavidin Dynabeads magnetic beads can be used to target biotinylated DNA fragments, followed by denaturation of the double-stranded DNA and removal of the non-biotinylated strand. The streptavidin-coupled Dynabeads magnetic beads will not inhibit any enzymatic activity. This enables further handling and manipulation of the bead-bound DNA directly on the solid phase. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/preparing-single-stranded-dna-templates.html) for more information in regards to single-stranded DNA capture.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

What is the magnetic susceptibility for Dynabeads magnetic beads?

Magnetic susceptibility is a measure of how quickly the beads will migrate to the magnet. This will depend on the iron content and the character of the iron oxide. The magnetic susceptibility given for the Dynabeads magnetic beads is the mass susceptibility, given either as cgs units/g or m^3/kg (the latter being an SI unit). For ferri- and ferromagnetic substances, the magnetic mass susceptibility is dependent upon the magnetic field strength (H), as the magnetization of such substances is not a linear function of H but approaches a saturation value with increasing field. For that reason, the magnetic mass susceptibility of the Dynabeads magnetic beads is determined by a standardized procedure under fixed conditions. The magnetic mass susceptibility given in our catalog is thus the SI unit. Conversion from Gaussian (cgs, emu) units into SI units for magnetic mass susceptibility is achieved by multiplying the Gaussian factor (emu/g or cgs/g) by 4 pi x 10^-3. The resulting unit is also called the rationalized magnetic mass susceptibility, which should be distinguished from the (SI) dimensionless magnetic susceptibility unit. In general, magnetic mass susceptibility is a measure of the force (Fz) influencing an object positioned in a nonhomogenous magnetic field. The magnetic mass susceptibility of the Dynabeads magnetic beads is measured by weighing a sample, and then subjecting the sample to a magnetic field of known strength. The weight (F1) is then measured, and compared to the weight of the sample when the magnetic field is turned off (F0). The susceptibility is then calculated as K x 10^-3 = [(F1-F0) x m x 0.335 x 10^6], where K is the mass susceptibility of the sample of mass m. The susceptibility is then converted to SI units.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

How can I determine coupling efficiency of Dynabeads magnetic beads?

There are different methods to check binding of ligands to the beads, including optical density (OD) measurement, fluorescent labeling, and radioactive labeling.

For OD measurement, you would measure the OD of the ligand before immobilization to the beads and compare it with the ligand concentration that is left in the supernatant after coating. This gives a crude measurement of how much protein has bound to the beads.

Protocol:

1.Set spectrophotometer to the right wavelength. As a blank, use the Coupling Buffer.
2.Measure the absorbance of the Pre-Coupling Solution. A further dilution may be necessary to read the absorbance, depending upon the amount of ligand added.
3.Measure the absorbance of the Post-Coupling Solution. A dilution may be necessary to read the absorbance.
4.Calculate the coupling efficiency, expressed as the % protein uptake, as follows. [(Pre-Coupling Solution x D) - (Post-Coupling Solution x D)] x 100/(Pre-Coupling Solution x D) where D = dilution factor.

For fluorescent labeling, we suggest negatively quantifying the amount of ligand bound by measuring ligand remaining in the coupling supernatant (compared to the original sample), rather than directly measuring the ligands on the beads. Add labeled ligand to the beads, and measure how much ligand is left in the supernatant (not bound to the beads). By comparing this with the total amount added in the first place, you can then calculate how much of the ligand that has been bound to the beads. Keep in mind that the Dynabeads magnetic beads are also autofluorescent, which is why direct measuring of fluorescence of the bead-bound ligands is not recommended, but rather this indirect approach. The label could be, for example, FITC/PE. Some researchers perform a direct approach with success (using a flow cytometer).

Radioactive labeling is the most sensitive method of the three, but it is also the most difficult one. It involves radioactively labeling a portion of the ligand. We use radiolabeled I-125 in tracer amounts and mix it with "cold" ligands in a known ratio before coupling. The absolute quantities for the ligand on the beads should be obtained by measuring the beads in a scintillation (gamma) counter and comparing the cpm with a standard.

Protocol:

1.Take out an appropriate amount of beads and wash the beads in 1 mL of binding buffer.
2.Pipette out desired amount of human IgG in a separate tube.
3.Mix the human IgG with I-125-labeled human IgG (30,000 - 100,000 cpm).
4.Dilute the mixture of human IgG and I-125-labeled human IgG to 100 mL in binding buffer.
5.Incubate for 30 minutes at room temperature and measure the cpm in a scintillation counter.
6.Wash the beads (with coating) four times, and measure cpm again.
The % binding is calculated by using the equation : (cpm after washing/cpm before washing)x100%.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.