Purified Mouse Anti-Human CD40
Clone 5C3 (RUO)
- Brand BD Pharmingen™
- Alternative Name TNFRSF5; TNF receptor superfamily member 5; CD40L receptor; Bp50; p50
- Concentration 125 µg/ml
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing) Rhesus, Cynomolgus, Baboon (Tested in Development)
Flow cytometry (Routinely Tested)
Immunohistochemistry-frozen, Immunohistochemistry-zinc-fixed (Tested During Development)
Immunohistochemistry-paraffin (Not Recommended)
- Workshop No. V CD40.4
- Entrez Gene ID 958
- Storage Buffer Aqueous buffered solution containing BSA, goat serum, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Suggested Companion Products
Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Immunohistochemistry: The 5C3 antibody is recommended to test for immunohistochemical staining of acetone-fixed frozen sections. Tissued tested were human spleen and tonsil. The antibody stains tonsillar B lymphocytes and some lymphoid dendritic cells. The isotype control recommended for use with this antibody is Purified Mouse IgG1 κ Isotype Control (Cat. No. 550878). For optimal indirect immunohistochemical staining, the 5C3 antibody should be titrated (1:10 to 1:50 dilution) and visualized via a three-step staining procedure in combination with polyclonal, biotin conjugated anti-mouse Igs (multiple adsorbed) (Cat. No. 550337) as the secondary antibody and Streptravidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880). A detailed protocol of the immunohistochemical procedure is available under "Cell Biology (WB, IP, IHC, IF)" at our website, http://www.bdbiosciences.com/us/s/resources. The clone 5C3 is not recommended for formalin-fixed paraffin embedded sections.