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R718 Mouse Anti-Human CD137
R718 Mouse Anti-Human CD137
Flow cytometric analysis of CD137 expression on stimulated human peripheral blood lymphocytes. Concanavalin A (Con-A)-stimulated (3 days) peripheral blood mononuclear cells were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD137 antibody (Cat. No. 567086/567223; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing the expressed levels of CD137 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) activated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD137 expression on stimulated human peripheral blood lymphocytes. Concanavalin A (Con-A)-stimulated (3 days) peripheral blood mononuclear cells were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD137 antibody (Cat. No. 567086/567223; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing the expressed levels of CD137 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) activated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
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BD Horizon™
4-1BB, ILA, TNFRSF9
Human (QC Testing)
Mouse BALB/c IgG1, κ
Recombinant Human 4-1BB Fusion Protein
Flow cytometry (Routinely Tested)
5 µl
VI C-7
3604
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567223 Rev. 7
Antibody Details
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4B4-1

The 4B4-1 monoclonal antibody specifically binds to CD137 which is also known as 4-1BB, and ILA (induced by lymphocyte activation). CD137 is a type I transmembrane glycoprotein that belongs to the TNF/NGF receptor family. It is encoded by TNFRSF9 (tumor necrosis factor receptor superfamily, member 9). CD137 is expressed on activated T cells, B cells, monocytes, and follicular dendritic cells. CD137 plays roles in the costimulation, differentiation and survival of T cells and B cells.

The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567223 Rev. 7
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567223 Rev.7
Citations & References
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Development References (6)

  1. Garni-Wagner BA, Lee ZH, Kim YJ, Wilde C, Kang CY, Kwon BS. 4-1BB is expressed on CD45RAhiROhi transitional T cell in humans. Cell Immunol. 1996; 169(1):91-98. (Immunogen: ELISA, Flow cytometry, Immunoprecipitation). View Reference
  2. Goodwin, RG. CDw137 (4-1BB) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:883-884.
  3. Hurtado JC, Kim SH, Pollok KE, Lee ZH, Kwon BS. Potential role of 4-1BB in T cell activation. Comparison with the costimulatory molecule CD28. J Immunol. 1995; 155(7):3360-3367. (Biology). View Reference
  4. Kim YJ, Broxmeyer HE. Therapeutic potential of 4-1BB (CD137) as a regulator for effector CD8(+) T cells. J Hematother Stem Cell Res. 2001; 10(4):441-449. (Biology). View Reference
  5. Zacharakis N, Chinnasamy H, Black M, et al. Immune recognition of somatic mutations leading to complete durable regression in metastatic breast cancer. Nat Med. 2018; 24(6):724-730. (Clone-specific: Flow cytometry). View Reference
  6. Zhou Z, Kim S, Hurtado J, et al. Characterization of human homologue of 4-1BB and its ligand. Immunol Lett. 1995; 45(1-2):67-73. (Biology). View Reference
View All (6) View Less
567223 Rev. 7

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.