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Cell-Vive™ GMP CD137 (4-1BB) Activation Beads

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S18014C.Rec (See other available formats)
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1_CV_BEADS_CD137_41BB_Activation_030626
Human peripheral mononuclear cells were isolated from whole blood and then stimulated with either the Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads or with Cell-Vive™ GMP CD137 (4-1BB) Activation Beads for 14 days. Cells were cultured in NKCCM using the suggested protocol. After 14 days, cells were harvested and analyzed for CD56+ fold expansion.
  • 1_CV_BEADS_CD137_41BB_Activation_030626
    Human peripheral mononuclear cells were isolated from whole blood and then stimulated with either the Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads or with Cell-Vive™ GMP CD137 (4-1BB) Activation Beads for 14 days. Cells were cultured in NKCCM using the suggested protocol. After 14 days, cells were harvested and analyzed for CD56+ fold expansion.
  • 2_CV_BEADS_CD137_41BB_Activation_030326
    Human peripheral mononuclear cells were isolated from whole blood and CD56+ NK cells were isolated using the MojoSort™ Human NK Cell Isolation Kit (cat#: 480054). Isolated NK cells were then stimulated with a combination of the Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads and the Cell-Vive™ GMP CD137 (4-1BB) Activation Beads at a 1:3 ratio for 11 days. Cells were cultured in NKCCM using the suggested protocol above. After 11 days, the beads were removed from culture using a magnetic separator, and the cells were re-stimulated with either the combined beads again at a 1:3 ratio (black bars) or the Cell-Vive™ GMP 4-1BB/CD137 Activation Beads alone (purple bars). Cells were re-stimulated for a total of 14 days and analyzed for (A) total fold expansion over D0 and (B) phenotype by flow cytometry.
  • 3_CV_BEADS_CD137_41BB_Activation_030626
  • 4_CV_BEADS_CD137_41BB_Activation_030326
    Human peripheral mononuclear cells were isolated from whole blood and then stimulated with the Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads. Cells were cultured for 11 days using the suggested protocol . On D11, cells were harvested, beads were removed on a magnetic separator, and cells were re-stimulated with either the Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads alone (black bars), the Cell-Vive™ GMP CD137 (4-1BB) Activation Beads alone (purple bars), or a combination of the Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads and the Cell Vive™ GMP CD137 (4-1BB) Activation Beads at a 1:1 ratio (grey bars) for an additional 7 days. At D7, cells were harvested for (A) total fold expansion over D0, (B) viability at D7 of re-stimulation, and (C) phenotype by flow cytometry.
  • 5_CV_BEADS_CD137_41BB_Activation_030326
  • 6_CV_BEADS_CD137_41BB_Activation_030626
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422613 0.5 mL 348€
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422614 2.5 mL 988€
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Description

Cell Vive™ GMP CD137 (4-1BB) Activation Beads allow for convenient in vitro or ex vivo activation of human lymphocytes without the need for antigen-presenting cells or feeder cells. Human T cells or NK cells are stimulated with anti-4-1BB antibodies coated on superparamagnetic microbeads. Simply add the beads directly to isolated T cells, isolated NK cells, or to PBMCs at a desired bead-to-cell ratio for robust activation or re-stimulation.

 

The Cell-Vive™ GMP 4-1BB/CD137 Activation Beads can be used in combination with Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads to achieve higher CD3+/CD8+ cell yields than when using CD3/CD28 stimulation alone. This combinatorial approach allows for flexibility in antibody-to-cell ratios that allow for optimized expansion when using different cell sources, donors, or clones of a particular cell type. When used for re-stimulation, the Cell Vive™ GMP CD137 (4-1BB) Activation Beads lead to higher CD8+ percentages over the use of CD3/CD28 for re-stimulation, regardless of the original source of T cell activation. The use of the Cell Vive™ GMP CD137 (4-1BB) Activation Beads for T cell re-stimulation also leads to higher fold expansion, higher viability, and reduced expression of exhaustion markers when compared to cells re-stimulated with CD3/CD28. We see this phenotype regardless of the initial activation condition or bead-to-cell ratio used.

 

When used for NK cell expansion and culture, the Cell Vive™ GMP CD137 (4-1BB) Activation Beads can be used for initial NK cell activation and expansion with similar results as the Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads. When used for re-stimulation, the Cell Vive™ GMP CD137 (4-1BB) Activation Beads alone provide similar expansion when compared to combinatorial approaches.

 

Excess antibody-coated beads can be removed from the cell suspension using a magnetic separator (some residual beads may be retained in the cell suspension). Some of the downstream applications include functional assays, gene expression analysis, and phenotypic characterization.

Product Details
Quality Statement

BioLegend Cell-Vive™ GMP Activation and Expansion Tools are manufactured and tested in accordance with USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue Engineered Products and Ph. Eur. Chapter 5.2.12 in a dedicated GMP facility compliant with ISO 13485:2016. Specifications and processes include

  • Low endotoxin level (< 1 EU/mL)
  • Manufactured under serum-free conditions
  • Bioburden/sterility and mycoplasma testing
  • Batch-to-batch consistency
  • Vendor qualification
  • Raw material traceability and documentation
  • Documented procedures and employee training
  • Equipment maintenance and monitoring records
  • Lot-specific certificates of analysis
  • Quality audits per ISO 13485:2016
  • QA review of released products
Technical Data Sheet (pdf)

Product Details

Reactivity
Human
Formulation
Beads are resuspended in phosphate buffer solution containing 0.1% recombinant HSA.
Endotoxin Level
< 1 EU/mL
Preparation
The antibodies were purified by affinity chromatography and conjugated with magnetic beads under optimized conditions. The final product is a GMP, ready-to-use solution.
Concentration
4.0 x 107 ± 10 % beads/mL
Storage & Handling
4°C (2°C-8°C)
Application

FC - Quality tested

Activ, Cell Culture - Verified in house
Recommended Usage

Cell Vive™ GMP CD137 (4-1BB) Activation Beads have been tested at different bead-to-cell ratios to validate different experimental conditions. Working concentrations should be obtained by titrating the beads for specific culture protocols and applications. Please see suggested protocols for further details.

Application Notes

Cell Vive™ GMP CD137 (4-1BB) Activation Beads are designed for the in vitro or ex vivo expansion of human T cells and NK cells. This product is a ready to use solution, and there is no need to dilute or reconstitute the product prior to use in cell culture.

 

Cell Vive™ GMP CD137 (4-1BB) Activation Beads are designed to assist in the activation and expansion of human T cells or NK cells, in tandem with other cell activation methods or alone. When used alone, this product is best utilized for re-stimulation.

 

This product has been optimized on human PBMCs, isolated human CD3+ T cells, and isolated CD56+ NK cells from whole blood. A suggested protocol for these cell types is below, but this protocol will require optimization depending on the intended use and cell source.

 

Suggested T cell expansion protocol in tandem with Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads (cat# 422605):

  1. Prepare desired T cell complete media (TCCM) using sterile, filtered cell-culture grade reagents. Standard TCCM formulation: IMDM with 5% Cell-Vive™ T-NK Xeno-Free Serum Substitute, GMP (cat#: 420502) and 200 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925).
  2. Prepare cells at a concentration of 1 x 106 per mL of TCCM.
  3. Prepare the activation bead mixture of Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads and Cell-Vive™ GMP 4-1BB/CD137 Activation Beads by mixing desired volume of beads in a 5 mL polypropylene tube with 1 mL of TCCM.
    1. When using PBMCs, ratios of CD3/CD28 beads and 4-1BB beads of 1:1, 1:3, and 1:4 have shown similar levels of fold expansion and final CD3+/CD8+ percentages. The optimal ratio of beads will depend on the cell source, media formulation, and cell culture scale.
    2. If using a 1:1 bead-to-cell ratio and a 1:1 CD3/CD28-to-4-1BB bead ratio, mix 12.5 µL of each bead type for a final bead mixture volume of 25 µL.
  4. Wash the beads by applying the tube with beads to a magnet (cat# 480019) for 1- 2 minutes. Decant supernatant and remove tube from the magnet.
  5. Resuspend the beads in 1 mL of cell suspension and plate in a 12 well cell culture plate.
  6. Allow cells to expand at 37°C and 5% CO2. At D4 and D7, count cells and re-seed at 0.5 x 106 cells per mL.
  7. At D10 or D11, harvest cells for desired applications or re-stimulation.

 

Suggested NK cell expansion protocol in tandem with Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads (cat# 422611):

  1. Prepare desired NK cell complete media (NKCCM) using sterile, filtered cell-culture grade reagents. A suggested NKCCM formulation is: IMDM with 5% human AB serum or serum substitute, 100 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925), and 15 ng/mL of Cell-Vive™ GMP Recombinant Human IL-15 (carrier-free) (cat#: 570314).
  2. Prepare cells at a concentration of 1 x 106 per mL of TCCM.
  3. Prepare the activation bead mixture of Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads and Cell-Vive™ GMP 4-1BB/CD137 Activation Beads by mixing desired volume of beads in a 5 mL polypropylene tube with 1 mL of NKCCM.
    1. When using PBMCs, a ratio of NKp46/CD2 beads and 4-1BB beads of 1:1 and 1:3 have shown optimal levels of fold expansion. The optimal ratio of beads will depend on the cell source, media formulation, and cell culture scale.
    2. If using a 1:1 bead-to-cell ratio and a 1:3 NKp46/CD2-to-4-1BB bead ratio, mix 6.25 µL of NKp46/CD2 beads and 18.75 µL of 4-1BB beads for a final bead mixture volume of 25 µL.
  4. Wash the beads by applying the tube with beads to a magnet (cat# 480019) for 1-2 minutes. Decant supernatant and remove tube from the magnet.
  5. Resuspend the beads in 1 mL of cell suspension and plate in a 12 well cell culture plate.
  6. Allow cells to expand at 37°C and 5% CO2. At D6, D9, and D12, count cells and re-seed at 1.0 x 106 cells per mL.
  7. At D14, harvest cells for desired applications or re-stimulation.

 

Suggested re-stimulation protocol for T cells or NK cells:

  1. Prepare desired cell culture media using sterile, filtered cell-culture grade reagents.
    1. A suggested NK cell complete media (NKCCM) formulation is: IMDM with 5% human AB serum or serum substitute, 100 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925), and 15 ng/mL of Cell-Vive™ GMP Recombinant Human IL-15 (carrier-free) (cat#: 570314).
    2. A suggested TCCM formulation is: IMDM with 5% Cell-Vive™ T-NK Xeno-Free Serum Substitute, GMP (cat#: 420502) and 200 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925).
  2. Prepare cells at a concentration of 1 x 106 per mL of media.
  3. Prepare the Cell-Vive™ GMP 4-1BB/CD137 Activation Beads for wash by mixing 25 µL. of beads in a 5 mL polypropylene tube with 1 mL of media.
    1. This volume will provide a 1:1 bead-to-cell ratio for re-stimulation. The optimal bead-to-cell ratio should be determined experimentally for each cell culture protocol and cell type.
  4. Wash the beads by applying the tube with beads to a magnet for 1-2 minutes. Decant supernatant and remove tube from the magnet.
  5. Resuspend the beads in 1 mL of cell suspension and plate in a 12 well cell culture plate.
  6. Allow cells to expand at 37°C and 5% CO2.
    1. For T cells, count cells on D4 and D7. Re-seed at 0.5 x 106 cells per mL.
    2. For NK cells, count cells on D6, D9, and D12. Re-seed at 1.0 x 106 cells per mL.
  7. At desired endpoint, harvest cells for desired applications and analysis.
Disclaimer

BioLegend Cell-Vive™ GMP Activation and Expansion tools are for research use only. Suitable for ex vivo cell processing. Not for use in diagnostic or therapeutic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

Antigen Details

Antigen References
  1. Gruss HJ & Dower SK. 1995. Blood. 85:3378-404.
  2. Sica G & Chen L. 2000. Adv Exp Med Biol. 465:355-62.
  3. Alderson MR, et al. 1994. Eur J Immunol. 24:2219-27.
  4. Schwarz H, et al. 1996. Blood. 87:2839-45.
Gene ID
NA

Related FAQs

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Other Formats

View All 4-1BB Reagents
Description Clone Applications
Cell-Vive™ GMP CD137 (4-1BB) Activation Beads S18014C.Rec FC,Cell Culture,Activ
Go To Top Version: 1    Revision Date: 03/19/2026

For Research Use Only. Suitable for ex vivo cell processing use. Not for injection or diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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