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R718 Mouse Anti-Human CD40
R718 Mouse Anti-Human CD40

Multiparameter flow cytometric analysis of CD40 expression on Human peripheral blood leukocyte populations.  Platelet-depleted whole blood was first treated with BD FACS Lysing™ Solution (Cat. No. 349202) to lyse erythrocytes. After washing, the leukocytes were then stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 567470; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD40 antibody (Cat. No. 569848/569924; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD40 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Multiparameter flow cytometric analysis of CD40 expression on Human peripheral blood leukocyte populations.  Platelet-depleted whole blood was first treated with BD FACS Lysing™ Solution (Cat. No. 349202) to lyse erythrocytes. After washing, the leukocytes were then stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 567470; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD40 antibody (Cat. No. 569848/569924; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD40 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Product Details
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BD Horizon™
CD40; TNFRSF5; Bp50
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Reported)
Mouse IgG1, κ
Human CD40
Flow cytometry (Routinely Tested)
5 µl/test
V B CD40.5; VIII 377
958
AB_3685368
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. 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.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  9. For U.S. patents that may apply, see bd.com/patents.
569924 Rev. 1
Antibody Details
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HB14

This HB14 monoclonal antibody specifically recognizes CD40 which is also known as Tumor necrosis factor receptor superfamily member 5 (TNFRSF5). CD40 is an ~45-48 kDa single-pass type I transmembrane glycoprotein. CD40 is expressed on B lymphocytes, monocytes, macrophages, dendritic cells, endothelial cells, fibroblasts, keratinocytes, and CD34+ hematopoietic progenitor cells. CD40 serves as a receptor for CD154 (CD40 Ligand or CD40L). CD40:CD154 interactions costimulate B cell growth, differentiation, and immunoglobulin isotype switching in antibody responses to T-dependent antigens as well as the generation of memory B cells and cytokine production by monocytes and dendritic cells. The HB14 antibody reportedly crossreacts with CD40 expressed by several non-human primates including Rhesus and Cynomolgus Macaques as well as Olive Baboons.

569924 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
R718
Red 627-640 nm
695 nm
718 nm
569924 Rev.1
Citations & References
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View product citations for antibody "569924" on CiteAb

Development References (12)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. Bjornson-Hooper ZB, Fragiadakis GK, Spitzer MH, et al. A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates.. Front Immunol. 2022; 13:867015. (Clone-specific: Flow cytometry). View Reference
  3. Clark EA, Ledbetter JA. Activation of human B cells mediated through two distinct cell surface differentiation antigens, Bp35 and Bp50. Proc Natl Acad Sci U S A. 1986; 83(12):4494-4498. (Biology). View Reference
  4. Freedman JE. CD40-CD40L and platelet function: beyond hemostasis.. Circ Res. 2003; 92(9):944-6. (Biology). View Reference
  5. Galy AH, Spits H. CD40 is functionally expressed on human thymic epithelial cells. J Immunol. 1992; 149(3):775-782. (Biology). View Reference
  6. Hu HM, O'Rourke K, Boguski MS, Dixit VM. A novel RING finger protein interacts with the cytoplasmic domain of CD40.. J Biol Chem. 1994; 269(48):30069-72. (Clone-specific: Immunoprecipitation). View Reference
  7. Inwald DP, McDowall A, Peters MJ, Callard RE, Klein NJ. CD40 is constitutively expressed on platelets and provides a novel mechanism for platelet activation.. Circ Res. 2003; 92(9):1041-8. (Biology). View Reference
  8. Itoh N, Yonehara S, Ishii A, et al. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell. 1991; 66(2):233-243. (Biology). View Reference
  9. Kansas GS, Tedder TF. Transmembrane signals generated through MHC class II, CD19, CD20, CD39, and CD40 antigens induce LFA-1-dependent and independent adhesion in human B cells through a tyrosine kinase-dependent pathway. J Immunol. 1991; 147(12):4094-4102. (Immunogen: Functional assay). View Reference
  10. Mitchell K, Barreyro L, Todorova TI, et al. IL1RAP potentiates multiple oncogenic signaling pathways in AML.. J Exp Med. 2018; 215(6):1709-1727. (Clone-specific: Flow cytometry). View Reference
  11. Pound JD, Challa A, Holder MJ, et al. Minimal cross-linking and epitope requirements for CD40-dependent suppression of apoptosis contrast with those for promotion of the cell cycle and homotypic adhesions in human B cells.. Int Immunol. 1999; 11(1):11-20. (Clone-specific: Functional assay). View Reference
  12. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
View All (12) View Less
569924 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.

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.