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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.
BD Pharmingen™ APC Hamster Anti-Mouse CD137 (4-1BB)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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.
Development References (3)
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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
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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
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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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.