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Flow cytometric analysis of CD134 expression on stimulated human peripheral blood lymphocytes. Phytohemagglutinin-stimulated (PHA; Sigma L-1668; 3 days) peripheral blood mononuclear cells were stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD134 antibody (Cat. No. 568213; solid line histogram) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing CD134 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV421 Mouse Anti-Human CD134
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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
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead 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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The ACT35 monoclonal antibody specifically binds to CD134 which is also known as OX40. The 35 kDa CD134 polypeptide is encoded by the TNFRSF4 gene. CD134 is a type I integral membrane glycoprotein and member of the tumor necrosis factor/nerve growth factor receptor (TNFR/NGFR) family. CD134 is expressed on activated T lymphocytes, hematopoietic precursor cells and fibroblasts. CD134 functions as a T cell costimulatory receptor when bound by OX40 Ligand/TNFSF4 that is expressed by antigen presenting cells. CD134 thereby plays roles in T-cell activation as well as the regulation of differentiation, proliferation or apoptosis of normal and malignant lymphoid cells. Analysis of the nucleotide sequence of the human TNFRSF4 cDNA reveals strong homology with the rat Tnfrsf4 cDNA sequence. OX40 was clustered as CD134 in the Sixth International Workshop on Human Leukocyte Differentiation Antigens.
Development References (7)
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Harrop JA, Spampinato J, Reddy M, Eichman C, Cook R, Truneh A. CD134 Workshop: Kinetics of expression of tumor necrosis factor receptor molecules and other cytokine receptors on activated CD4-positive cells. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:871-873.
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Latza U, Durkop H, Schnittger S, et al. The human OX40 homolog: cDNA structure, expression and chromosomal assignment of the ACT35 antigen. Eur J Immunol. 1994; 24(3):677-683. (Clone-specific: Western blot). View Reference
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Merl A, Pohla H, Adibzadeh M, Paelec G. CD13r Workshop: Expression of cytokine receptors on anergic CD4-positive TCR2-positive TH0 cell clones. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:873-875.
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Moffat S, Higashimura N, Sugamura K. CD134 (OX40) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:869-871.
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Schwarting R, Stein H. ACT35: a new mAb recognizing a 35-kDa antigen with a tissue distribution similar to that of the CD25 molecule (interleukin-2 receptor). In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:464-465.
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Schwarting R, Stein H. Report on single/unclustered and provisionally grouped antibodies. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:460-463.
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