Flow cytometric analysis of CD262 (TRAIL-R2) expression Panel 1. Mouse NIH/3T3 cells. Cells from the mouse NIH/3T3 (Embryonic Fibroblast, ATCC CRL-1658) cell line were stained with PE Hamster IgG2, κ Isotype Control (Cat. No. 550085; dashed line histogram) or PE Hamster Anti-CD262 (TRAIL-R2) antibody (Cat. No. 566813; solid line histogram) at 0.5 μg/test. The fluorescence histogram showing CD262 (TRAIL-R2) expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable cells. Panel 2. Activated mouse splenic T lymphocytes. Concanavalin A-stimulated (3 days) BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 (Cat. No. 561091/553051; Plots 2a and 2b) and FITC Rat Anti-Mouse CD8a (Cat. No. 561966/553030/553031; Plots 2c and 2d) antibodies, and either PE Hamster IgG2, κ Isotype Control (Plots 2a and 2c) or PE Hamster Anti-CD262 (TRAIL-R2) antibody (Plots 2b and 2d) at 0.5 μg/test. Nonviable cells were excluded from the analysis by staining with DAPI (Cat. No. 564907). Two-color flow cytometric contour plots showing the correlated expression of CD4 or CD8 versus CD262 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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Flow cytometric analysis of CD262 (TRAIL-R2) expression Panel 1. Mouse NIH/3T3 cells. Cells from the mouse NIH/3T3 (Embryonic Fibroblast, ATCC CRL-1658) cell line were stained with PE Hamster IgG2, κ Isotype Control (Cat. No. 550085; dashed line histogram) or PE Hamster Anti-CD262 (TRAIL-R2) antibody (Cat. No. 566813; solid line histogram) at 0.5 μg/test. The fluorescence histogram showing CD262 (TRAIL-R2) expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable cells. Panel 2. Activated mouse splenic T lymphocytes. Concanavalin A-stimulated (3 days) BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 (Cat. No. 561091/553051; Plots 2a and 2b) and FITC Rat Anti-Mouse CD8a (Cat. No. 561966/553030/553031; Plots 2c and 2d) antibodies, and either PE Hamster IgG2, κ Isotype Control (Plots 2a and 2c) or PE Hamster Anti-CD262 (TRAIL-R2) antibody (Plots 2b and 2d) at 0.5 μg/test. Nonviable cells were excluded from the analysis by staining with DAPI (Cat. No. 564907). Two-color flow cytometric contour plots showing the correlated expression of CD4 or CD8 versus CD262 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
Tnfrsf10b; TRAILR2; DR5; TRICK2; KILLER; Ly98; MK
Mouse (QC Testing)
Armenian Hamster IgG, κ
Mouse DR5 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869881
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE Hamster IgG2, κ Isotype Control RUO
Size
0.1 mg
Cat No.
550085
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566813 Rev. 1
Antibody Details
MD5-1
The MD5-1 monoclonal antibody specifically recognizes CD262 which is also known as TNF-related apoptosis inducing ligand receptor 2 (TRAIL-R2/TRAILR2), TRICK2, Death receptor 5 (DR5), KILLER, MK, or Ly98. CD262 (TRAIL-R2) is a ~55 kDa type I transmembrane protein that is encoded by Tnfrsf10b (tumor necrosis factor receptor superfamily, member 10b). The extracellular region of this receptor contains three TNFR-cysteine repeats which is followed by a transmembrane region and a cytoplasmic region with a death domain. CD262 (TRAIL-R2) forms a homotrimeric complex that is expressed on activated T cells, B cells, and plasma cells as well as some tumors and tumor cell lines. TRAIL (CD253) binds to CD262 (TRAIL-R2) which leads to the association of its cytoplasmic death domain with FADD and other factors leading to activation of the NF-κB pathway and cellular apoptosis. The complex interplay of TRAIL with CD262 (TRAIL-R2) and other TRAIL receptors contributes to the regulation of immunity and prevention of autoimmunity as well as host defense against cancer and regulation of bone resorption. The MD5-1 antibody can reportedly induce TRAIL-mediated apoptosis of TRAIL-sensitive target cells within in vitro and in vivo experimental model systems.
566813 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566813 Rev.1
Citations & References
View product citations for antibody "566813" on CiteAb
Development References (9)
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Condamine T, Kumar V, Ramachandran IR, et al. ER stress regulates myeloid-derived suppressor cell fate through TRAIL-R-mediated apoptosis.. J Clin Invest. 2014; 124(6):2626-39. (Clone-specific: Flow cytometry, Functional assay, In vivo exacerbation). View Reference
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Degli-Esposti M. To die or not to die—the quest of the TRAIL receptors. J Leukoc Biol. 1999; 65:535-542. (Biology).
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Dufour F, Rattier T, Shirley S, et al. N-glycosylation of mouse TRAIL-R and human TRAIL-R1 enhances TRAIL-induced death.. Cell Death Differ. 2017; 24(3):500-510. (Clone-specific: Flow cytometry, Functional assay). View Reference
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LeBlanc HN, Ashkenazi A. Apo2L/TRAIL and its death and decoy receptors. Cell Death Differ. 2003; 10:66-75. (Biology).
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Schneider P, Olson D, Tardivel A, et al. Identification of a new murine tumor necrosis factor receptor locus that contains two novel murine receptors for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). J Biol Chem. 2003; 278:5444-5454. (Biology).
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Smyth MJ, Takeda K, Hayakawa Y, Peschon JJ, van den Brink MR, Yagita H. Nature's TRAIL—on a path to cancer immunotherapy. Immunity. 2003; 18:1-6. (Biology).
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Takeda K, Yamaguchi N, Akiba H, et al. Induction of tumor-specific T cell immunity by anti-DR5 antibody therapy.. J Exp Med. 2004; 199(4):437-48. (Immunogen: Flow cytometry, Functional assay). View Reference
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Ursini-Siegel J, Zhang W, Altmeyer A, et al. TRAIL/Apo-2 ligand induces primary plasma cell apoptosis. J Immunol. 2002; 169:5505-5513. (Biology).
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Wu GS, Burns TF, Zhan Y, Alnemri ES, El-Deiry WS. Molecular cloning and functional analysis of the mouse homologue of the KILLER/DR5 tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor. Cancer Res. 1999; 59:2770-2775. (Biology).
566813 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.