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

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

Product No.: L322

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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
<1.0 EU/µg as determined by the LAL method
Purity
≥95% 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.
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)

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The TKS-1 clone is used in in vivo mouse studies primarily for targeting the mouse 4-1BB ligand (4-1BBL), also known as CD137L. This ligand is crucial in immune responses, particularly in the activation and survival of T cells, including CD8+ T cells, which are involved in antitumor immunity.

Role of 4-1BBL and TKS-1:

  • 4-1BBL (CD137L) is a type II transmembrane glycoprotein expressed on antigen-presenting cells (APCs) and can also be found on activated T cells. It interacts with 4-1BB (CD137) on T cells to enhance their activation and survival, which is vital for sustained immune responses against tumors.
  • TKS-1 Monoclonal Antibody is designed to react specifically with mouse 4-1BBL. By using TKS-1 in mouse studies, researchers can modulate the 4-1BB/4-1BBL pathway. This can either enhance or inhibit immune responses depending on the context of the study. For example, blocking the interaction might be used to study immune suppression or tolerance, while agonistic approaches could augment antitumor immunity by promoting T cell activation and proliferation.

In Vivo Applications:

  • Antitumor Studies: TKS-1 can be used to enhance antitumor immunity by promoting the activation and survival of CD8+ T cells, which are critical for tumor clearance.
  • Immune Regulation: By modulating the 4-1BB/4-1BBL pathway, researchers can study mechanisms of immune tolerance and autoimmunity, providing insights into how to balance immune responses to prevent autoimmune diseases while maintaining effective antitumor immunity.

Overall, the TKS-1 clone is a valuable tool in in vivo mouse studies for understanding and manipulating immune responses, particularly in the context of cancer research and immunotherapy.

The search results do not provide specific information about the storage temperature for a sterile packaged clone TKS-1. However, for general biological materials, especially DNA or bacteria, it is common to store them at -20°C to preserve their integrity and viability. For specific instructions on the storage of TKS-1, it would be best to consult the manufacturer's guidelines or the product documentation provided with the clone.

If TKS-1 is related to biological vectors similar to the pCold System, it might require storage at -20°C, similar to the pCold DNA vectors.

Commonly used antibodies or proteins alongside TKS-1 in the literature include co-stimulatory and checkpoint molecules, as well as T-cell markers and peptide antigens involved in immune response modulation.

Key antibodies and proteins cited with TKS-1:

  • 3H3: A monoclonal anti-4-1BB antibody. It is frequently used in combination with TKS-1 to investigate their synergistic effects on T-cell activation, proliferation, and tumor suppression.
  • CD8 (CD8?, CD8?): Used to identify and quantify CD8^+ T cells, which are a major subset analyzed in immunological studies using TKS-1.
  • CD62L and CCR7: Surface markers indicative of T-cell activation and migration status, often measured alongside TKS-1 treatment to assess T-cell differentiation and trafficking.
  • Sphingosine-1 phosphate receptor 1 (S1PR1): Another protein marker investigated for its upregulation after TKS-1 treatment, reflecting increased T-cell egress from lymphoid organs.
  • Thy1.1: An allelic marker used to track the transfer and proliferation of specific T-cell populations in adoptive transfer experiments with TKS-1.
  • mgp100 peptide: Antigenic peptide (mouse gp100) used to activate antigen-specific T cells, especially in the context of tumor immunity models with TKS-1.
  • IgG (isotype control antibodies): Employed as a negative control in studies to validate the specificity of TKS-1 effects.

In summary, studies employing TKS-1 commonly use antibodies or proteins such as 3H3, CD8, CD62L, CCR7, S1PR1, Thy1.1, and relevant antigenic peptides (e.g., mgp100), often in mouse models of T-cell activation, differentiation, and immunotherapy.

The key findings from scientific literature citing clone TKS-1 indicate that this antibody induces only mild agonistic signaling of 4-1BB compared to other clones, such as 3H3, which strongly triggers 4-1BB signals with minimal or no 4-1BBL signals. TKS-1 is used experimentally to study the function and signaling of 4-1BB (CD137) in immune cells, particularly in models assessing self-reactive CD8+ T cell survival and immune regulation.

  • TKS-1 shows mild agonist activity: When administered in vivo, TKS-1 produces modest activation of the 4-1BB pathway, which is less robust than that induced by clone 3H3. This characteristic makes it useful for dissecting the nuances of 4-1BB-mediated immune responses.

  • Role in cis-interaction studies: TKS-1 has been valuable in studying 4-1BB/4-1BBL cis-interactions on the surface of immune cells, helping delineate the contributions of these signals to cell survival, especially among self-reactive CD8+ T cells. Such investigations have expanded understanding of how immune cell fate decisions are regulated through receptor/ligand interactions.

  • Comparison to other clones: The contrast between TKS-1’s mild action and stronger agonists like 3H3 has enabled researchers to clarify the effects of differing signal strengths on immune outcomes.

These findings have contributed to immunological research, particularly in deciphering costimulatory signal modulation and the potential therapeutic manipulation of T cell responses. There is no evidence in indexed literature linking clone TKS-1 to replication controversies or citation impact debates relevant to nonreplicable publications.

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.