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APC Hamster Anti-Mouse CD137 (4-1BB)
APC Hamster Anti-Mouse CD137 (4-1BB)
Multicolor flow cytometric analysis of CD137 (4-1BB) expression on viable stimulated Mouse splenic leukocytes.  Concanavalin A (Con A)-activated (3 days) BALB/c Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The leukocytes were then stained with PE Rat Anti-Mouse CD25 antibody (Cat. No. 553075) and with either no APC conjugated antibody (Autofluorescence control; Left Plot) or APC Hamster Anti-Mouse CD137 (4-1BB) antibody (Cat. No. 570316/570394; Right Plot) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD137 (4-1BB) [or cellular Autofluorescence] versus CD25 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD137 (4-1BB) expression on viable stimulated Mouse splenic leukocytes.  Concanavalin A (Con A)-activated (3 days) BALB/c Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The leukocytes were then stained with PE Rat Anti-Mouse CD25 antibody (Cat. No. 553075) and with either no APC conjugated antibody (Autofluorescence control; Left Plot) or APC Hamster Anti-Mouse CD137 (4-1BB) antibody (Cat. No. 570316/570394; Right Plot) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD137 (4-1BB) [or cellular Autofluorescence] versus CD25 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
4-1BB; Cd137; ILA; Ly63; Tnfrsf9
Mouse (QC Testing)
Syrian Hamster IgG
Mouse 2PK-3 B Lymphoma Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
21942
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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 cell and BD® CompBeads 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. For U.S. patents that may apply, see bd.com/patents.
570316 Rev. 1
Antibody Details
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17B5

The 17B5 monoclonal antibody specifically recognizes CD137 which is also known as 4-1BB. CD137 (4-1BB) is an ~39 kDa single-pass type I transmembrane glycoprotein that is encoded by Tnfsf9 (Tnfsf9 tumor necrosis factor ligand superfamily, member 9). Monomeric and multimeric forms of CD137 (4-1BB) are expressed on thymocytes as well as activated T cells, B cells, NK cells, monocytes, macrophages, and dendritic cells (DC). It is also expressed on T regulatory cells and may function in their proliferation. CD137 (4-1BB) plays a costimulatory role in the interactions between T cells, B cells, and antigen presenting cells as it binds to 4-1BB ligand (4-1BBL), which is also known as CD137 ligand (CD137L). The 17B5 antibody can reportedly block these costimulatory interactions.

570316 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
570316 Rev.1
Citations & References
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View product citations for antibody "570316" on CiteAb

Development References (3)

  1. Zahm CD, Colluru VT, McNeel DG. Vaccination with High-Affinity Epitopes Impairs Antitumor Efficacy by Increasing PD-1 Expression on CD8(+) T Cells. Cancer Immunology Research. 2017; 5(8):630-641. (Clone-specific: Flow cytometry). View Reference
  2. Zahm CD, Moseman JE, Delmastro LE, G Mcneel D. PD-1 and LAG-3 blockade improve anti-tumor vaccine efficacy.. Oncoimmunology. 2021; 10(1):1912892. (Clone-specific: Flow cytometry). View Reference
  3. Zheng G, Wang B, Chen A. The 4-1BB costimulation augments the proliferation of CD4+CD25+ regulatory T cells.. J Immunol. 2004; 173(4):2428-34. (Clone-specific: Blocking, Flow cytometry, Functional assay). View Reference
570316 Rev. 1

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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.