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General description
Tumor necrosis factor receptor superfamily member 5 (UniProt: P25942; also known as B-cell surface antigen CD40, Bp50, CD40L receptor, CDw40, CD40) is encoded by the CD40 (also known as TNFRSF5) gene (Gene ID: 958) in human. CD40, a member of the TNFR super family, is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-20 ), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 21-193), a transmembrane domain (aa 194-215), and a cytoplasmic domain (aa 216-277). It has multiple cysteine-rich repeats. CD40 is expressed in B-cells, monocytes, dendritic cells, thymic epithelium as well as on certain carcinomas. It serves as a receptor for CD40L that is transiently expressed on T-cells and some non-immune cells under inflammatory conditions. Upon engagement with its ligand, CD40 directly or indirectly recruits TRAF1, TRAF2, TRAF3, TRAF5, and TRAF6 to its cytoplasmic domains. The canonical consensus binding sites for TRAF1, TRAF2, and TRAF3 are situated at the membrane distal domain of the intracellular tail of CD40 and is defined by the amino acid sequence PxQxT. CD40 transduces TRAF6 and MAP3K8-mediated signals that activate ERK in macrophages and B cells, leading to induction of immunoglobulin secretion. Clone G28.5 is reported to bind to an epitope localized in D1/B2 domains in the extracellular cysteine rich region and is a potent activator of B-cells. Mutations in CD40 gene are known to cause immunodeficiency with hyper-IgM3 (HIGM3) that is characterized by normal or elevated serum IgM levels with absence of IgG, IgA, and IgE. This results in a profound susceptibility to bacterial infections. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Elgueta, R., et al. (2009). Immunol. Rev. 229(1); 152-172; Malmborg Hager, AC., et al. (2003). Scand. J, Immunol. 57(6); 517-524).
We are committed to bringing you greener alternative products, which adhere to one or more of the 12 Principles of Green Chemistry. This antibody is preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed, and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 20 amino acids from the N-terminal half of human CD40.
Application
Quality Control Testing
Evaluated by Western Blotting in Raji cell lysate.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected CD40 in Raji cell lysate.
Tested Applications
Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected CD40 in lysates from THP-1 cells treated with LPS and Human tonsil tissue, and with His/ABP-tagged recombinant fragment of human CD40
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected CD40 in Human tonsil tissue sections.
Enzyme Immunoassay (ELISA): A serial of dilutions from a representative lot detected His/ABP-tagged recombinant fragment of human CD40
Affinity Binding Assay: A representative lot of this antibody bound CD40 peptide with a KD of 7.6 x 10-9 in an affinity binding assay.
Flow Cytometry Analysis: 1 µg from a representative lot detected CD40 in one million Human peripheral blood mononuclear cells (PBMC) co-stained with CD19APC.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 2E20 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects CD40. It targets an epitope within 20 amino acids within the extracellular domain from the N-terminal half.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Segment | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 2E20, recombinant monoclonal |
| description | recombinant, expressed in HEK 293 cells |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt 30.62 kDa,observed mol wt ~38 kDa |
| purified by | using Protein A |
| species reactivity | human |
| packaging | antibody small pack of 25 |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| technique(s) | ELISA: suitable,affinity binding assay: suitable,flow cytometry: suitable,immunohistochemistry: suitable,western blot: suitable |
| isotype | IgG |
| epitope sequence | Extracellular domain |
| Protein ID accession no. | NP_001241 |
| UniProt accession no. | P25942 |
| greener alternative category | Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
| Gene Information | human ... CD40(958) |
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