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Purified NA/LE Mouse Anti-Human CD40L (CD154)
Purified NA/LE Mouse Anti-Human CD40L (CD154)
Flow cytometric analysis of 24-31 antibody-mediated inhibition of CD25 upregulation by CD3-stimulated Human PBMC.    Panel A. PBMC were cultured with plate-bound Purified NA/LE Mouse anti-Human CD3 antibody (Cat. No. 555336; 18 h) and with 1 µg of either Purified NA/LE Mouse IgG1, κ Isotype Control (Cat. No. 554721/553447; Upper Plot) or Purified NA/LE Mouse Anti-Human CD40L (CD154) antibody (Cat. No. 568010; Lower Plot). The cells were washed and preincubated with Human BD Fc Block™ (Cat. No. 564219/564220), and then stained with PE Mouse anti-Human CD25 antibody (Cat. No. 555432/557138) and APC Mouse anti-Human CD19 antibody (Cat. No. 555415/561742). DAPI Solution (Cat. No. 564907) was added to cells right before analysis. Contour plots showing the expression of CD25 by CD19 positive or CD19 negative cells were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes.    Panel B. PBMC were similarly stimulated in the absence (0 µg; dashed line histogram) or presence of the indicated doses (0.05-2 µg) of the Purified NA/LE Isotype Control (dotted line histograms) or Purified NA/LE Anti-Human CD40L (CD154) antibody (solid line histograms). The cultured cells were stained as previously described. Histograms showing CD25 expression were derived from CD19 positive-gated viable lymphocytes.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of 24-31 antibody-mediated inhibition of CD25 upregulation by CD3-stimulated Human PBMC.    Panel A. PBMC were cultured with plate-bound Purified NA/LE Mouse anti-Human CD3 antibody (Cat. No. 555336; 18 h) and with 1 µg of either Purified NA/LE Mouse IgG1, κ Isotype Control (Cat. No. 554721/553447; Upper Plot) or Purified NA/LE Mouse Anti-Human CD40L (CD154) antibody (Cat. No. 568010; Lower Plot). The cells were washed and preincubated with Human BD Fc Block™ (Cat. No. 564219/564220), and then stained with PE Mouse anti-Human CD25 antibody (Cat. No. 555432/557138) and APC Mouse anti-Human CD19 antibody (Cat. No. 555415/561742). DAPI Solution (Cat. No. 564907) was added to cells right before analysis. Contour plots showing the expression of CD25 by CD19 positive or CD19 negative cells were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes.    Panel B. PBMC were similarly stimulated in the absence (0 µg; dashed line histogram) or presence of the indicated doses (0.05-2 µg) of the Purified NA/LE Isotype Control (dotted line histograms) or Purified NA/LE Anti-Human CD40L (CD154) antibody (solid line histograms). The cultured cells were stained as previously described. Histograms showing CD25 expression were derived from CD19 positive-gated viable lymphocytes.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
CD40 ligand; CD40-L; CD40LG; T-BAM; TNFSF5; TRAP; gp39; hCD40L
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human gp39 Protein
Blocking, Flow cytometry (Tested During Development)
1.0 mg/ml
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  4. An isotype control should be used at the same concentration as the antibody of interest.
568010 Rev. 2
Antibody Details
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24-31

The 24-31 monoclonal antibody specifically binds to CD40 Ligand (CD40L) which is also known as CD154, gp39, T-B cell-activating molecule (T-BAM), Tumor necrosis factor ligand superfamily member 5 (TNFSF5), and TNF-related activation protein (TRAP). CD40L (CD154) is an ~39 kDa type II membrane glycoprotein that is encoded by CD40LG. It is expressed on a variety of cell types including activated CD4+ T cells and some CD8+ T cells, NK cells, mast cells and basophils. CD40L (CD154) serves as a ligand for CD40 that is expressed on B cells, macrophages, and dendritic cells.  The expression of CD40L (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 CD40L (CD154) and CD40 interaction is required for B-lymphocyte differentiation, including immunoglobulin production and isotype switching and memory B cell generation. The 24-31 antibody can reportedly 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 CD40L (CD154) due to mutations in the CD40LG gene.

568010 Rev. 2
Format Details
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NA/LE
NA/LE refers to the culture and purification methods and buffer used to produce purified antibodies with no azide and low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.NA/LE are perfectly suited to be used in culture or in vivo (for nonhuman studies) for functional assays — blocking, neutralizing, activation or depletion — where the presence of azide may damage cells or exogenous endotoxin may signal or activate cells.
NA/LE
568010 Rev.2
Citations & References
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View product citations for antibody "568010" on CiteAb

Development References (4)

  1. Brams P, Black A, Padlan EA, et al. A humanized anti-human CD154 monoclonal antibody blocks CD154-CD40 mediated human B cell activation.. Int Immunopharmacol. 2001; 1(2):277-94. (Clone-specific: Blocking). View Reference
  2. Foy TM, McIlraith M, Masters SR, et al. Blockade of CD40-CD154 interferes with human T cell engraftment in scid mice.. Cell Transplant. 7(1):25-35. (Immunogen: Blocking). View Reference
  3. 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
  4. 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). View Reference
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568010 Rev. 2

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