Anti-Mouse CD137L (4-1BBL) – Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD137L (4-1BBL) – Purified in vivo PLATINUM™ Functional Grade

Product No.: L320

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Clone
TKS-1
Target
4-1BBL
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
4-1BBL, CD137L, TNFSF9, 4-1BB Ligand, TKS-1
Isotype
Rat IgG2a κ
Applications
FC
,
in vivo

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
BALB/c mouse B lymphoma line 2PK-3
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
Next Day 2-8°C
Additional Applications Reported In Literature ?
FC
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
TKS-1 activity is directed against 4-1BBL (CD137L).
Background
4-1BBL (CD137L) and 4-1BB (CD137) are a receptor ligand pair belonging to the tumor necrosis factor (TNF) receptor/TNF superfamily1,2. 4-1BBL is a type II transmembrane protein expressed by splenic B and T cells, macrophages, and dendritic cells1-5. Members of the TNF-TNF receptor superfamily play critical roles in regulating cellular activation, differentiation, and apoptosis6 and the 4-1BBL-4-1BB interaction is important in cellular immune responses5. 4-1BBL-4-1BB interaction provides a co-stimulatory signal to T cells, resulting in increased proliferation and cytokine production5.

A TKS-1-producing hybridoma was generated by immunizing a SD rat with 4-1BBL-transfected NRK cells (rat IgG2a,κ)5. Splenocytes were fused with P3U1 cells and hypoxanthine–aminopterin–thymidine selection was performed. The monoclonal antibody was identified by its strong reactivity with 4-1BBL-transfected L5178Y cells, cloned by limiting diffusion, purified from ascites, and found to bind to 4-1BBL/NRK and 4-1BBL/P815 cells5.

Pre-incubation with TKS-1 blocks 4-1BB–Ig binding to 4-1BBL/L5178Y cells, indicating TKS-1 is specific to mouse 4-1BBL and can interrupt the interaction between 4-1BBL and 4-1BB5. Additionally, TKS-1 can block anti-CD3-stimulated T cell proliferation co-stimulated by 4-1BBL /P815 cells. The monoclonal antibodies 19H3 and TKS-1 bind to different sites on murine 4-1BBL, with TKS-1 binding to a site that overlaps with the receptor binding site7. Additionally, TKS-1 and 4-1BB bind to a similar site, making TKS-1 especially useful for blocking studies.
Antigen Distribution
4-1BBL is expressed by activated B cells, macrophages, and dendritic cells.
Ligand/Receptor
4-1BB (CDw137)

Leinco Antibody Advisor

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The most common in vivo application of clone TKS-1 in mice is to block or study the function of mouse 4-1BB ligand (4-1BBL, also called CD137L), a critical costimulatory molecule involved in T cell activation and survival, particularly of CD8+ T cells in immune and antitumor responses.

Key in vivo applications of TKS-1 in mice include:

  • Blocking the 4-1BB–4-1BBL interaction: TKS-1 specifically binds to mouse 4-1BBL and can block its interaction with the 4-1BB receptor on T cells. This interaction is essential for delivering a costimulatory signal that augments T cell proliferation, cytokine secretion, and survival.

  • Functional studies of immune regulation: By blocking the 4-1BB–4-1BBL pathway, TKS-1 is used to study the role of this costimulatory signal in disease models such as infection, cancer, or autoimmunity—typically by evaluating changes in T cell responses, proliferation, memory formation, and antitumor immunity.

  • Antitumor immune response experiments: Since 4-1BBL supports the activation and persistence of cytotoxic T lymphocytes, TKS-1 can be administered in vivo to dissect the contribution of 4-1BB signaling to the effectiveness of immunotherapy or tumor rejection models.

  • Blocking T cell costimulation in adoptive transfer or vaccine models: TKS-1 has been used to inhibit costimulatory signals during experiments involving T cell transfer or antigen-specific immune activation, allowing researchers to clarify the dependence of these responses on 4-1BB–4-1BBL signaling.

TKS-1 is well-validated for in vivo use in blocking studies and mechanistic immunological assays in mice, making it a critical reagent for dissecting the functional role of 4-1BBL in T cell biology and antitumor immunity.

Commonly Used Proteins and Antibodies Alongside TKS-1

TKS-1 is most commonly referenced as a monoclonal antibody targeting mouse 4-1BBL (CD137L), a member of the tumor necrosis factor receptor superfamily. In the literature, TKS-1 is often used in combination with other antibodies or proteins to investigate immunological mechanisms, particularly in the context of T-cell activation, migration, and antitumor responses.

Directly Associated Antibodies and Proteins

  • 3H3 Antibody: In several studies, TKS-1 is used in combination with the 3H3 antibody, which targets 4-1BB (CD137). The co-administration of 3H3 (anti-4-1BB) and TKS-1 (anti-4-1BBL) has been shown to modulate the survival, proliferation, and migration of antigen-specific CD8+ T cells in mouse models, with the combination being more effective than either antibody alone in certain contexts.
  • 17B5 Antibody: This is mentioned as a control antibody in some experiments alongside TKS-1 and 3H3, though its precise target is not specified in the provided literature.
  • Mouse gp100 (mgp100) Peptide: Used to stimulate and track pmel-1 Thy1.1+ CD8+ T cells in vivo, this peptide is a model antigen in studies involving TKS-1 to assess T-cell responses.

Broader Context: Other Antibodies and Targets in Similar Research

While not specifically used with TKS-1 in the provided literature, it is common in immunological research to encounter antibodies against:

  • CD8, CD4, CD62L, and CCR7 for T-cell subset and activation/migration analysis.
  • Sphingosine-1 phosphate receptor 1 (S1PR1) for investigating T-cell egress from lymphoid organs, which is referenced in the context of TKS-1’s effects on T-cell migration.
  • Control antibodies such as isotype-matched IgGs are routinely used as negative controls in these types of studies.

Potential Mechanisms and Pathways

  • TCR Signaling: The combination of 3H3 and TKS-1 is proposed to amplify TCR signaling, enhancing T-cell division and differentiation.
  • Type I IFN-related response: Transcriptome analysis in the presence of TKS-1 shows upregulation of genes involved in interferon response, T-cell activation, proliferation, and migration.

Summary Table

NameTargetRole/Application with TKS-1Reference
3H34-1BB (CD137)Co-stimulation of T cells, combined with TKS-1 for enhanced effects
17B5Not specifiedControl antibody in experimental setups
mgp100Model antigenStimulates pmel-1 CD8+ T cells
CD62L/CCR7Homing receptorsAssess T-cell migration upon TKS-1 treatment
S1PR1Sphingosine receptorInvestigate T-cell egress from lymphoid organs

Key Points

  • TKS-1 is most frequently paired with 3H3 (anti-4-1BB) in the literature to study synergistic effects on T-cell responses.
  • Mouse gp100 peptide is used as a model antigen to trace antigen-specific T-cell responses in these experiments.
  • Control antibodies and migration/homing markers (CD62L, CCR7, S1PR1) are commonly analyzed alongside TKS-1 to dissect immunological mechanisms.
  • Other antibodies or proteins are not explicitly mentioned in the provided literature as directly used with TKS-1, but the above represent the core toolkit in the cited studies.

If you have a specific experimental context or disease model in mind, additional antibodies or proteins may be relevant, but based on the available literature, 3H3 and mgp100 are the most directly associated with TKS-1 in published studies.

Key findings from scientific literature citing clone TKS-1 focus on its specificity, functional properties, and applications in immunology research, particularly in the context of mouse 4-1BB ligand (4-1BBL/CD137L) biology.

  • Specificity and Mechanism: TKS-1 is a monoclonal antibody that binds specifically to mouse 4-1BBL, blocking the interaction between 4-1BBL and its receptor 4-1BB. It can interrupt the co-stimulatory signaling that 4-1BBL provides to T cells, thus modulating immune cell activation.

  • Epitope and Binding Site: TKS-1 binds to a site on murine 4-1BBL that overlaps with the natural 4-1BB receptor binding site, making it especially effective for blocking studies. It is often contrasted with other clones, like 19H3, which bind to different epitopes. This property enables TKS-1 to uniquely disrupt ligand-receptor engagement and is valuable in mechanistic studies of the 4-1BB/4-1BBL axis in immune responses.

  • Functional Effects Compared to Other Clones: Unlike the 3H3 clone, which induces strong agonistic signaling through 4-1BB, TKS-1 triggers only mild agonistic effects. This makes TKS-1 more suited for blocking or neutralizing studies rather than for driving robust receptor activation. When co-administered with 3H3, TKS-1 can attenuate some effects of strong 4-1BB activation (e.g., decreasing total lymph node cell numbers in some models).

  • Applications: TKS-1 is widely used in animal (mouse) models to investigate the role of 4-1BBL in T cell activation, proliferation, and cytokine production. It has been instrumental in delineating the involvement of 4-1BBL in immune regulation by facilitating selective blockade of the 4-1BB/4-1BBL interaction.

  • Research Utility: Due to its blocking ability and epitope specificity, TKS-1 is a tool for:

    • Defining the functional contribution of 4-1BBL in immune responses
    • Dissecting the relative roles of different co-stimulatory pathways in mouse models
    • Differentiating between receptor- and ligand-mediated signaling in experimental settings

In summary, literature citations of TKS-1 emphasize its high specificity for mouse 4-1BBL, its capacity to block receptor-ligand interaction, and its mild agonist activity relative to other clones, making it a preferred blocking antibody for mechanistic studies of 4-1BB/4-1BBL signaling in murine immunology.

There is currently no published standard dosing regimen specific for clone TKS-1 (anti-mouse CD137L/4-1BBL) across different mouse models. According to product information resources and available literature, dosing protocols for TKS-1 have not been systematically reported or standardized in the scientific literature as of the present.

Key context:

  • TKS-1 is an anti-mouse CD137 ligand (4-1BBL) antibody, often used to modulate immune responses in experimental settings, but unlike other well-characterized monoclonal antibodies, its dosing regimens have not been broadly detailed in peer-reviewed studies or product datasheets in a way that supports cross-model comparison.
  • Related antibody-based regimens (for other targets) are typically determined empirically based on mouse strain, disease model, study objectives, and intended immunomodulatory effect. These regimens can include variables such as dose amount (e.g., mg/kg), frequency (single vs. multiple injections), and route of administration (i.p., i.v., s.c.), but the specifics for TKS-1 are not reported.
  • Investigators often select dosing regimens based on pilot studies, or extrapolate from similar anti-CD137L antibodies or related immune-targeting antibodies when direct guidance is lacking.

Implications:

  • If you are planning an experiment involving clone TKS-1 in mice, dosing should be determined based on experimental design, pilot titration, and consultation with published studies using similar CD137L-targeting antibodies or by contacting suppliers/experts directly.
  • For rigorous studies, reporting complete details of dosing regimen—including amount, frequency, administration route, and animal model details—is critical for reproducibility in future research.

In summary:
There is no established or widely published dosing regimen for clone TKS-1 across different mouse models. Experimental variables, research aims, and pilot optimization will drive dosing choices in the absence of published standards.

References & Citations

1. Goodwin RG, Din WS, Davis-Smith T, et al. Eur. J. Immunol. 23:2631-2641. 1993.
2. Alderson MR, Smith CA, Tough TW, et al. Eur. J. Immunol. 24:2219-2227. 1994.
3. Pollok KE, Kim YJ, Hurtado J, et al. Eur J Immunol. 24(2):367-374. 1994.
4. DeBenedette MA, Shahinian A, Mak TW, et al. J. Immunol. 158:551-559. 1997.
5. Futagawa T, Akiba H, Kodama T, et al. Int Immunol. 14(3):275-286. 2002.
6. Smith CA, Farrah T, Goodwin RG. Cell 76:959-962. 1994.
7. Mbanwi AN, Lin GHY, Wang KC, et al. J Immunol Methods. 450:81-89. 2017.
8. Zheng G, Wang B, Chen A. J Immunol. 173(4):2428-2434. 2004.
9. Madireddi S, Eun SY, Lee SW, et al. J Exp Med. 211(7):1433-1448. 2014.
10. Shrestha S, Noh JM, Kim SY, et al. Oncoimmunology. 5(1):e1067744. 2015.
11. Kang SW, Lee SC, Park SH, et al. Cancer Res. 77(21):5989-6000. 2017.
Flow Cytometry
in vivo Protocol

Certificate of Analysis

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Formats Available

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.