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V450 Mouse Anti-Human CD40
V450 Mouse Anti-Human CD40

Flow cytometric analysis of CD40 expression on human peripheral blood  lymphocytes. Whole blood was stained with BD Horizon™ V450 Mouse Anti-Human CD40 antibody (Cat. No. 561219; solid line histogram) or with a BD Horizon™ V450 Mouse IgG1, κ Isotype Control (Cat. No. 560373; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD40 expression on human peripheral blood  lymphocytes. Whole blood was stained with BD Horizon™ V450 Mouse Anti-Human CD40 antibody (Cat. No. 561219; solid line histogram) or with a BD Horizon™ V450 Mouse IgG1, κ Isotype Control (Cat. No. 560373; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
TNFRSF5; TNF receptor superfamily member 5; CD40L receptor; Bp50; p50
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
V CD40.4
958
AB_10715579
Aqueous buffered solution containing protein stabilizer, glycerol 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 with BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 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. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561219 Rev. 3
Antibody Details
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5C3

This 5C3 monoclonal antibody specifically binds to CD40, a 45-48 kDa type I integral membrane glycoprotein. CD40 is expressed on B lymphocytes, but is not expressed on terminally differentiated B cells. CD40 is also expressed by endothelial cells, basal epithelial cells and some epithelial cell carcinomas, follicular dendritic cells, macrophages, fibroblasts, keratinocytes, and CD34+ hematopoietic progenitor cells. This antibody is useful for studying the roles played by CD40 in B-cell growth, proliferation, and differentiation including immunoglobulin isotype switching. Anti-CD40 antibodies have been reported to stimulate B-cell proliferation when costimulated with anti-µ, anti-CD20 antibodies or with phorbol esters. 5C3 is capable of inducing B-cell proliferation when presented with IL-4.

Clone 5C3 reacts with the human form of the 45-48 kDa type I integral membrane glycoprotein, CD40. This clone also cross-reacts with a subset of peripheral blood lymphocytes, but not monocytes nor granulocytes, of baboon and both rhesus and cynomolgus macaque monkeys. The distribution on lymphocytes is similar to that seen with normal human donor lymphocytes, with the reactivity being restricted to CD20+ lymphocytes.

The antibody is conjugated to BD Horizon V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon V450 can be used in place of Pacific Blue™ conjugates.

561219 Rev. 3
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
V450
Violet 405 nm
405 nm
450 nm
561219 Rev.3
Citations & References
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Development References (5)

  1. Morio T, Hanissian SH, Bacharier LB, et al. Ku in the cytoplasm associates with CD40 in human B cells and translocates into the nucleus following incubation with IL-4 and anti-CD40 mAb. Immunity. 1999; 11(3):339-348. (Biology). View Reference
  2. Nguyen LT, Duncan GS, Mirtsos C, et al. TRAF2 deficiency results in hyperactivity of certain TNFR1 signals and impairment of CD40-mediated responses. Immunity. 1999; 11(3):379-389. (Biology). View Reference
  3. Randall TD, Heath AW, Santos-Argumedo L, Howard MC, Weissman IL, Lund FE. Arrest of B lymphocyte terminal differentiation by CD40 signaling: mechanism for lack of antibody-secreting cells in germinal centers. Immunity. 1998; 8(6):733-742. (Biology). View Reference
  4. 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.
  5. Stamenkovic I, Clark EA, Seed B. A B-lymphocyte activation molecule related to the nerve growth factor receptor and induced by cytokines in carcinomas. EMBO J. 1989; 8(5):1403-1410. (Biology). View Reference
View All (5) View Less
561219 Rev. 3

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.