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Alexa Fluor® 647 Mouse Anti-Human CD258 (LIGHT)
Alexa Fluor® 647 Mouse Anti-Human CD258 (LIGHT)

Two-color flow cytometric analysis of CD258 (LIGHT) expression on stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated (36 hr) with Phorbol 12-Myristate 13-Acetate (PMA; Sigma-Aldrich, Cat. No. P-8139; 1 ng/ml) and Ionomycin (Sigma Cat. No. I-0634; 1 μg/ml). The cells were then stained with Alexa Fluor® 488 Mouse Anti-Human CD8 antibody (Cat. No. 557696/ 557704) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human CD258 (LIGHT) antibody (Cat. No. 564374; Right Panel). The two-color flow cytometric contour plots showing the correlated expression patterns of CD8 versus CD258 (LIGHT) [or Ig Isotype control staining] were generated for gated events with the forward and side-light scattering characteristics of viable activated lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometry System.

Two-color flow cytometric analysis of CD258 (LIGHT) expression on stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated (36 hr) with Phorbol 12-Myristate 13-Acetate (PMA; Sigma-Aldrich, Cat. No. P-8139; 1 ng/ml) and Ionomycin (Sigma Cat. No. I-0634; 1 μg/ml). The cells were then stained with Alexa Fluor® 488 Mouse Anti-Human CD8 antibody (Cat. No. 557696/ 557704) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human CD258 (LIGHT) antibody (Cat. No. 564374; Right Panel). The two-color flow cytometric contour plots showing the correlated expression patterns of CD8 versus CD258 (LIGHT) [or Ig Isotype control staining] were generated for gated events with the forward and side-light scattering characteristics of viable activated lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometry System.

Product Details
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BD Pharmingen™
TNFSF14; LIGHT; HVEML; LTg
Human (QC Testing)
Mouse IgG1, κ
Human LIGHT Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
IX
AB_2738775
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
564374 Rev. 1
Antibody Details
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115520

The 115520 monoclonal antibody specifically binds to CD258 which is also known as LIGHT (is homologous to lymphotoxins, exhibits inducible expression, and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes). CD258 is also referred to as HVEM-L (Herpes virus entry mediator ligand), and LT-γ. This type II transmembrane glycoprotein belongs to TNF superfamily; it is encoded by TNFSF14 (tumor necrosis factor ligand superfamily member 14). CD258 is expressed by activated T cells and cultured dendritic cells. CD258 can costimulate T cells and antigen presenting cells as well as trigger apoptosis by some cell types. CD258 binds to CD270/LIGHT-R/HVEM and to the Lymphotoxin β Receptor (LTβR). LIGHT reportedly also binds to the decoy receptor (DcR3/TR6).

564374 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564374 Rev.1
Citations & References
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Development References (3)

  1. Anderson DM, Maraskovsky E, Billingsley WL, et al. A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature. 1997; 390(6656):175-179. (Biology). View Reference
  2. Cannon JG, Kraj B, Sloan G. Follicle-stimulating hormone promotes RANK expression on human monocytes. Cytokine. 2011; 53(2):141-144. (Biology). View Reference
  3. Riegel A, Maurer T, Prior B, et al. Human polymorphonuclear neutrophils express RANK and are activated by its ligand, RANKL. Eur J Immunol. 2012; 42(4):975-981. (Biology). View Reference
564374 Rev. 1

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.