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562846_Image_2.png

Two-color flow cytometric analysis of CD40 expression on mouse splenocytes. Mouse splenic leucocytes were stained with FITC Rat Anti-Mouse CD4  (Cat. No. 553047/553046/561835) and FITC Rat Anti-Mouse CD8a (Cat. No. 553031/553030/561966), and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon™ BV421 Rat Anti-Mouse CD40 antibody (Cat. No. 562846; Right Panel). Two-color flow cytometric dot plots show the correlated expression of CD4 + CD8 versus CD40 (or Ig Isotype Control staining) for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

562846Image1.png

Two-color flow cytometric analysis of CD40 expression on mouse splenocytes. Mouse splenic leucocytes were stained with FITC Rat Anti-Mouse CD4  (Cat. No. 553047/553046/561835) and FITC Rat Anti-Mouse CD8a (Cat. No. 553031/553030/561966), and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon™ BV421 Rat Anti-Mouse CD40 antibody (Cat. No. 562846; Right Panel). Two-color flow cytometric dot plots show the correlated expression of CD4 + CD8 versus CD40 (or Ig Isotype Control staining) for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

562846_Image_2.png 562846Image1.png Blue-Cap-Violet-Label.png

Two-color flow cytometric analysis of CD40 expression on mouse splenocytes. Mouse splenic leucocytes were stained with FITC Rat Anti-Mouse CD4  (Cat. No. 553047/553046/561835) and FITC Rat Anti-Mouse CD8a (Cat. No. 553031/553030/561966), and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon™ BV421 Rat Anti-Mouse CD40 antibody (Cat. No. 562846; Right Panel). Two-color flow cytometric dot plots show the correlated expression of CD4 + CD8 versus CD40 (or Ig Isotype Control staining) for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Two-color flow cytometric analysis of CD40 expression on mouse splenocytes. Mouse splenic leucocytes were stained with FITC Rat Anti-Mouse CD4  (Cat. No. 553047/553046/561835) and FITC Rat Anti-Mouse CD8a (Cat. No. 553031/553030/561966), and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon™ BV421 Rat Anti-Mouse CD40 antibody (Cat. No. 562846; Right Panel). Two-color flow cytometric dot plots show the correlated expression of CD4 + CD8 versus CD40 (or Ig Isotype Control staining) for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
Bp50; Tnfrsf5; TNR5; TRAP; CD40L receptor; GP39; HIGM1; IMD3; T-BAM
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse CD40 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2734767
Aqueous buffered solution containing BSA 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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
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Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
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562846 Rev. 1
Antibody Details
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3/23

The 3/23 clone monoclonal antibody specifically binds to CD40, a 40-50 kDa glycoprotein expressed on B lymphocytes and other antigen-presenting cells.  CD40 has been reported to be transiently expressed on activated CD4+ and CD8+ T cells and in some mouse strains, the 3/23 mAb has been reported to react with 5-10% of T lymphocytes in adult mouse, but not neonatal, spleen.  CD40 plays a key role in B-cell growth and differentiation where interactions of CD40 with its ligand, CD154, are involved in the initiation, effector, and memory stages of cell-mediated immune responses.  In addition, CD40 has been reported to be involved with the triggering of NK cells and NK-T cells. Ligation of CD40 with the 3/23 antibody has been reported to induce splenic B cells to express the costimulatory molecule CD86 (B7-2).  In addition, although the 3/23 antibody by itself is a weak B-cell mitogen, it has been reported to synergize markedly with mitogenic anti-IgM, anti-IgD mAb or IL-4 to promote B-cell proliferation.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562846 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
562846 Rev.1
Citations & References
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View product citations for antibody "562846" on CiteAb

Development References (13)

  1. Bourgeois C, Rocha B, Tanchot C. A role for CD40 expression on CD8+ T cells in the generation of CD8+ T cell memory. Science. 2002; 297(5589):2060-2063. (Biology). View Reference
  2. Grewal IS, Flavell RA. CD40 and CD154 in cell-mediated immunity. Annu Rev Immunol. 1998; 16:111-135. (Biology). View Reference
  3. Hasbold J, Johnson-Leger C, Atkins CJ, Clark EA, Klaus GG. Properties of mouse CD40: cellular distribution of CD40 and B cell activation by monoclonal anti-mouse CD40 antibodies. Eur J Immunol. 1994; 24(8):1835-1842. (Immunogen: Activation, (Co)-stimulation). View Reference
  4. Hasbold J, Klaus GG. B cells from CBA/N mice do not proliferate following ligation of CD40. Eur J Immunol. 1994; 24(1):152-157. (Clone-specific: Activation, (Co)-stimulation). View Reference
  5. Klaus GG, Holman M, Hasbold J. Properties of mouse CD40: the role of homotypic adhesion in the activation of B cells via CD40. Eur J Immunol. 1994; 24(11):2714-2719. (Biology). View Reference
  6. Leifeld L, Trautwein C, Dumoulin FL, Manns MP, Sauerbruch T, Spengler U. Enhanced expression of CD80 (B7-1), CD86 (B7-2), and CD40 and their ligands CD28 and CD154 in fulminant hepatic failure. Am J Pathol. 1999; 154(6):1711-1720. (Biology: Immunohistochemistry). View Reference
  7. Mahnke K, Becher P, Ricciardi-Castagnoli P, Luger TA, Schawrz T Grabbe S. CD14 is expressed by subsets of murine dendritic cells and upregulated by lipopolysaccharide. In: Ricciardi-Castagnoli P, ed. Dendritic Cells in Fundamental and Clinical Immunology. New York: Plenum Press; 1997:145-159.
  8. Munder M, Mallo M, Eichmann K, Modolell M. Murine macrophages secrete interferon gamma upon combined stimulation with interleukin (IL)-12 and IL-18: A novel pathway of autocrine macrophage activation. J Exp Med. 1998; 187(12):2103-2108. (Biology). View Reference
  9. Noelle RJ, Ledbetter JA, Aruffo A. CD40 and its ligand, an essential ligand-receptor pair for thymus-dependent B-cell activation. Immunol Today. 1992; 13(11):431-433. (Biology). View Reference
  10. Parry SL, Hasbold J, Holman M, Klaus GG. Hypercross-linking surface IgM or IgD receptors on mature B cells induces apoptosis that is reversed by costimulation with IL-4 and anti-CD40. J Immunol. 1994; 152(6):2821-2829. (Biology). View Reference
  11. Parry SL, Holman MJ, Hasbold J, Klaus GG. Plastic-immobilized anti-mu or anti-delta antibodies induce apoptosis in mature murine B lymphocytes. Eur J Immunol. 1994; 24(4):974-979. (Biology). View Reference
  12. Tomura M, Yu WG, Ahn HJ, et al. A novel function of Valpha14+CD4+NKT cells: stimulation of IL-12 production by antigen-presenting cells in the innate immune system. J Immunol. 1999; 163(1):93-101. (Biology). View Reference
  13. Turner JG, Rakhmilevich AL, Burdelya L, et al. Anti-CD40 antibody induces antitumor and antimetastatic effects: the role of NK cells. J Immunol. 2001; 166(1):89-94. (Biology). View Reference
View All (13) View Less
562846 Rev. 1

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