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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 BD Horizon™ BV421 conjugated antibody (Autofluorescence control; Left Plot) or BD Horizon™ BV421 Hamster Anti-Mouse CD137 (4-1BB) antibody (Cat. No. 570317/570395; Right Plot) at 0.5 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 Horizon™ BV421 Hamster Anti-Mouse CD137 (4-1BB)
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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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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配套商品
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.
研发参考 (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). 查看参考
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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). 查看参考
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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). 查看参考
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