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BB515 Mouse Anti-Human CD40L (CD154)
BB515 Mouse Anti-Human CD40L (CD154)
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 BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dotted line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD40L (CD154) antibody (Cat. No. 568170/568171; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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 BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dotted line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD40L (CD154) antibody (Cat. No. 568170/568171; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
商品详情
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BD Horizon™
CD40 ligand; CD40-L; CD40LG; T-BAM; TNFSF5; TRAP; gp39; hCD40L
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human gp39 Protein
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


准备和存储

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 and unconjugated antibody and free dye were removed.

推荐的实验流程

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.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

商品通知

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. 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).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
抗体详情
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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.

格式详情
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
报价单和参考
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View product citations for antibody "568171" on CiteAb

研发参考 (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):277-294. (Clone-specific: Blocking, Flow cytometry). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
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