Flow cytometric analysis of LTβR expression by HeLa, Jurkat and human monocytes. HeLa cell line (Left Panel), Jurkat cell line (Middle Panel), or human peripheral monocytes (Right Panel) were stained with PE Mouse Anti-Human TNFR Related Protein (LTβR) (Cat No. 551503; filled histograms) or PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; empty histograms) at 0.5 µg/10^6 cells. Whole human blood was first treated with PharmLyse™ Lysing Buffer (Cat No. 555899) to lyse erythrocytes and blocked with normal polyclonal human IgG (5 µg/10^6 cells) prior to staining. Note that certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors. Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable cells.
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Flow cytometric analysis of LTβR expression by HeLa, Jurkat and human monocytes. HeLa cell line (Left Panel), Jurkat cell line (Middle Panel), or human peripheral monocytes (Right Panel) were stained with PE Mouse Anti-Human TNFR Related Protein (LTβR) (Cat No. 551503; filled histograms) or PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; empty histograms) at 0.5 µg/10^6 cells. Whole human blood was first treated with PharmLyse™ Lysing Buffer (Cat No. 555899) to lyse erythrocytes and blocked with normal polyclonal human IgG (5 µg/10^6 cells) prior to staining. Note that certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors. Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable cells.
Product Details
BD Pharmingen™
LT-BETA-R; LTBR; Lymphotoxin-beta receptor; TNF-RIII; TNFRSF3; TNFR-RP
Human (QC Testing)
Mouse IgG1, κ
Human LTβR-Fc protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_394222
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 www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
551503 Rev. 2
Antibody Details
hTNFR-RP-M12
The hTNFR-RP-M12 monoclonal antibody specifically recognizes the extracellular domain of the 61 kDa receptor for the human cytokines, LTα1β2 and LIGHT. This receptor is referred as the Lymphotoxin β Receptor (LTβR). LTβR is encoded by TNFRSF3. LTβR was previously known as TNFRIII and TNF receptor-related protein (TNFRrp). LTβR is a type I transmembrane glycoprotein and member of the TNF Receptor Superfamily. LTβR are expressed on stromal cells in lymphoid tissue, normal dermal fibroblasts, bronchial airway epithelial cells and in a variety of adherent cell lines including FDC-1, U937, HT-29, HeLa and HEK 293 cells. LTβR are absent on human peripheral blood T and B cells and expressed at low levels by monocytes.
551503 Rev. 2
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
551503 Rev.2
Citations & References
Development References (6)
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Browning JL, Dougas I, Ngam-ek A, et al. Characterization of surface lymphotoxin forms. Use of specific monoclonal antibodies and soluble receptors.. J Immunol. 1995; 154(1):33-46. (Immunogen). View Reference
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Crowe PD, VanArsdale TL, Walter BN, et al. A lymphotoxin-beta-specific receptor. Science. 1994; 264(5159):707-710. (Biology). View Reference
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Murphy M, Walter BN, Pike-Nobile L, . Expression of the lymphotoxin beta receptor on follicular stromal cells in human lymphoid tissues. Cell Death Differ. 1998; 5(6):497-505. (Biology). View Reference
-
Rooney IA, Butrovich KD, Glass AA, et al. The lymphotoxin-beta receptor is necessary and sufficient for LIGHT-mediated apoptosis of tumor cells. J Biol Chem. 2000; 275(19):14307-14315. (Biology). View Reference
-
Tamada K, Shimozaki K, Chapoval AI, . LIGHT, a TNF-like molecule, costimulates T cell proliferation and is required for dendritic cell-mediated allogeneic T cell response. J Immunol. 2000; 164(8):4105-4110. (Biology). View Reference
-
Zhai Y, Guo R, Hsu TL, et al. LIGHT, a novel ligand for lymphotoxin beta receptor and TR2/HVEM induces apoptosis and suppresses in vivo tumor formation via gene transfer.. J Clin Invest. 1998; 102(16):1142-1151. (Biology). View Reference
551503 Rev. 2
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.
