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Flow cytometric analysis of OX40 Ligand expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were preincubated with recombinant OX40 Ligand and stimulated with phytohemagglutinin (PHA; Sigma L-1668) for 3 days. The cells were then stained with either BD Horizon™ BV421 Mouse IgG1, k Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human OX40 Ligand (CD252) (Cat. No. 563766; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.


BD Horizon™ BV421 Mouse Anti-Human OX40 Ligand (CD252)

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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
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
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The ik-1 monoclonal antibody specifically binds to OX40 Ligand (OX40L) which is also known as CD252 and CD134 Ligand. OX40L is encoded by TNFSF4 (Tumor necrosis factor (ligand) superfamily, member 4). OX40L a 34 kDa type II transmembrane glycoprotein that is expressed by dendritic cells, B cells, human umbilical vein endothelial cells (HUVEC) and other types of vascular endothelial cells. OX40L serves as a costimulatory ligand by binding to and signaling through OX40 (CD134) which is expressed on activated T cells. It also mediates adhesion of activated T cells or OX40-positive transformed cells to vascular endothelial cells. Monocyte-derived dendritic cells (MoDC) express OX40 Ligand after stimulation through CD40. Ligation of OX40 Ligand expressed by activated MoDC can significantly increase their cytokine production.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

Development References (5)
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Imura A, Hori T, Imada K, et al. The human OX40/gp34 system directly mediates adhesion of activated T cells to vascular endothelial cells. J Exp Med. 1996; 183(5):2185-2195. (Biology). View Reference
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Kunitomi A, Hori T, Imura A, Uchiyama T. Vascular endothelial cells provide T cells with costimulatory signals via the OX40/gp34 system. J Leukoc Biol. 2000; 68(1):111-118. (Clone-specific: Blocking, Flow cytometry, Functional assay, Inhibition). View Reference
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Matsumura Y, Hori T, Kawamata S, Imura A, Uchiyama T. Intracellular signaling of gp34, the OX40 ligand: induction of c-jun and c-fos mRNA expression through gp34 upon binding of its receptor, OX40. J Immunol. 1999; 163(6):3007-3011. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition). View Reference
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Morimoto S, Kanno Y, Tanaka Y, et al. CD134L engagement enhances human B cell Ig production: CD154/CD40, CD70/CD27, and CD134/CD134L interactions coordinately regulate T cell-dependent B cell responses. J Immunol. 2000; 164(8):4097-4104. (Biology). View Reference
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Ohshima Y, Tanaka Y, Tozawa H, Takahashi Y, Maliszewski C, Delespesse G. Expression and function of OX40 ligand on human dendritic cells. J Immunol. 1997; 159(8):3838-3848. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.