-
Your selected country is
Norway
- Change country/language
Old Browser
This page has been recently translated and is available in French now.
Looks like you're visiting us from {countryName}.
Would you like to stay on the current country site or be switched to your country?
Flow cytometric analysis of CD40L (CD154) expression on stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMCs) were stimulated for 4 hours with 20 ng/mL Phorbol 12-Myristate 13-Acetate (PMA; Sigma-Aldrich Cat. No. P-8139) and 250 ng/mL Calcium Ionophore A23187 (Sigma-Aldrich Cat. No. C-9275). The cells were then preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 556648; dotted line histogram) or PE Mouse Anti-Human CD40L (CD154) antibody (Cat. No. 568007/568008; solid line histogram). DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing CD40L (CD154) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ PE Mouse Anti-Human CD40L (CD154)
Regulatory Status Legend
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 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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
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.
Development References (4)
-
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):277-294. (Clone-specific: Blocking). View Reference
-
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
-
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
-
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
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.