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PE-Cy™7 Mouse Anti-Human CD134
PE-Cy™7 Mouse Anti-Human CD134

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 PE-Cy™7 Mouse IgG1, κ Isotype Control (Cat. No. 557872; dashed line histogram) or PE-Cy™7 Mouse Anti-Human CD134 antibody (Cat. No. 563663; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

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 PE-Cy™7 Mouse IgG1, κ Isotype Control (Cat. No. 557872; dashed line histogram) or PE-Cy™7 Mouse Anti-Human CD134 antibody (Cat. No. 563663; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
OX40; TNFRSF4; TXGP1L; OX40L Receptor
Human (QC Testing)
Mouse IgG1, κ
Human HUT 102
Flow cytometry (Routinely Tested)
5 µl
IV A107, V BP048, V A068, VI C-31
7293
AB_2738358
Aqueous buffered solution containing BSA and ≤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 PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  6. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  7. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563663 Rev. 2
Antibody Details
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ACT35

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.

563663 Rev. 2
Format Details
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PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
563663 Rev.2
Citations & References
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Development References (7)

  1. 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.
  2. 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
  3. 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.
  4. 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.
  5. 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.
  6. 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.
  7. 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.
View All (7) View Less
563663 Rev. 2

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