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RY586 Mouse Anti-Human CD154
RY586 Mouse Anti-Human CD154
Flow cytometric analysis using BD OptiBuild™ RY586 Mouse Anti-Human CD154 antibody (Cat. No. 753194; solid line histogram) on human peripheral blood mononuclear stimulated with PMA and Ionomycin for 5 hours, with Isotype Control (Cat. No. 568097; dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Flow cytometric analysis using BD OptiBuild™ RY586 Mouse Anti-Human CD154 antibody (Cat. No. 753194; solid line histogram) on human peripheral blood mononuclear stimulated with PMA and Ionomycin for 5 hours, with Isotype Control (Cat. No. 568097; dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Product Details
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BD OptiBuild™
CD40LG; CD40 ligand; CD40L; TNFSF5; T-BAM; TRAP
Human (Tested in Development)
Mouse BALB/c IgG1, κ
Human TRAP1 Transfected Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
VI 6T-068
959
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Researchers should determine the optimal concentration of this reagent for their individual applications.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. CF™ is a trademark of Biotium, Inc.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  10. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
753194 Rev. 2
Antibody Details
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TRAP1

The TRAP1 monoclonal antibody specifically binds to CD154. CD154 is a 39 kDa type II membrane glycoprotein that is a member of the tumor necrosis factor superfamily, Tumor necrosis factor ligand superfamily member 5 (TNFSF5). CD154 is expressed on a variety of cell types including activated CD4+ T cells and some CD8+ T cells, NK cells, mast cells and basophils. CD154 is also known as CD40 ligand (CD40L); it serves as a ligand for CD40 that is expressed on B cells, macrophages, and dendritic cells.  The expression of CD154 by activated T-helper cells costimulates B-cell activation and proliferation through binding to CD40 expressed on B cells. In response to T-dependent antigens, the CD154 and CD40 interaction is required for B-lymphocyte differentiation, including immunoglobulin production and isotype switching and memory B cell generation. The TRAP1 antibody can partially block T cell-B cell interaction and inhibit the subsequent proliferation, differentiation, and memory formation of B cells. It has been reported that patients with X-linked hyper-IgM syndrome have defective expression of functional CD154 due to mutations in the CD40LG gene that encodes CD154.

753194 Rev. 2
Format Details
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RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RY586
Yellow-Green 561 nm
564 nm
586 nm
753194 Rev.2
Citations & References
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View product citations for antibody "753194" on CiteAb

Development References (10)

  1. Altenburg A, Baldus SE, Smola H, Pfister H, Hess S. CD40 ligand-CD40 interaction induces chemokines in cervical carcinoma cells in synergism with IFN-gamma. J Immunol. 1999; 162(7):4140-4147. (Clone-specific: Immunohistochemistry). View Reference
  2. Fuleihan R, Ramesh N, Horner A, et al. Cyclosporin A inhibits CD40 ligand expression in T lymphocytes. J Clin Invest. 1994; 93(3):1315-1320. (Biology). View Reference
  3. Gray D, Dullforce P, Jainandunsing S. Memory B cell development but not germinal center formation is impaired by in vivo blockade of CD40-CD40 ligand interaction. J Exp Med. 1994; 180(1):141-155. (Biology). View Reference
  4. 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.
  5. Kroczek RA, Graf D, Brugnoni D, et al. Defective expression of CD40 ligand on T cells causes "X-linked immunodeficiency with hyper-IgM (HIGM1)". Immunol Rev. 1994; 138:39-59. (Clone-specific: Flow cytometry). View Reference
  6. MacDonald KP, Nishioka Y, Lipsky PE, Thomas R. Functional CD40 ligand is expressed by T cells in rheumatoid arthritis. J Clin Invest. 1997; 100(9):2404-2414. (Clone-specific: Flow cytometry). View Reference
  7. Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002.
  8. Nishioka Y, Lipsky PE. The role of CD40-CD40 ligand interaction in human T cell-B cell collaboration. J Immunol. 1994; 153(3):1027-1036. (Biology). View Reference
  9. O'Gorman MR, Zaas D, Paniagua M, Corrochano V, Scholl PR, Pachman LM. Development of a rapid whole blood flow cytometry procedure for the diagnosis of X-linked hyper-IgM syndrome patients and carriers. Clin Immunol Immunopathol. 1997; 85(2):172-181. (Biology: Flow cytometry). View Reference
  10. van Kooten C, Gaillard C, Galizzi JP, et al. B cells regulate expression of CD40 ligand on activated T cells by lowering the mRNA level and through the release of soluble CD40. Eur J Immunol. 1994; 24(4):787-792. (Biology). View Reference
View All (10) View Less
753194 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.