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CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-81101

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Human

Applications

Block/Neutralize, ELISA, Functional, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Lambda Clone # A54 scFv

Format

Azide and BSA Free

Concentration

1 mg/ml

Product Specifications

Immunogen

A54 human anti-CD40/TNFRSF5 scFv was selected from the n-CoDeR phage display library after 4 rounds of selections using biotinylated CD40-Fcgamma fusion protein as bait and subsequent capture of bound phages on streptavidin-caoted magnetic beads.

Specificity

A54 binds specifically to the D1 domain of CD40/TNFRSF5 (and can only bind to the intact i.e. non-truncated version of CD40/TNFRSF5 and when an AD2 peptide tag has not been added) and can recognise CD40/TNFRSF5 on a B cell line (BJAB). This scFv binds to CD40/TNFRSF5 (immobilized by amine coupling) with a KD within the range of 40nM. CD40/TNFRSF5 is a glycoprotein of the TNFR superfamily and is expressed on all mature B cells, dendritic cells, activated monocytes, some endothelial cells and some epithelium including the thymus. CD40/TNFRSF5 binds to the membrane protein CD154 of the TNF superfamily and this interaction is required for the seconfary immune responses and germinal centre formation.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Lambda

Scientific Data Images for CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free

Immunocytochemistry/ Immunofluorescence: CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free [NBP2-81101]

Immunocytochemistry/ Immunofluorescence: CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free [NBP2-81101]

Immunocytochemistry/Immunofluorescence: CD40/TNFRSF5 Antibody (A54 scFv) [NBP2-81101] - Immunofluorescence staining of fixed Daudi cells with [NBP2-81101]. Immunofluorescence analysis of paraformaldehyde fixed Daudi cells on Shi-fix coverslips, permeabilized with 0.15% Triton and stained with the chimeric rabbit IgG version of A54 scFv [NBP2-81101] at 10 ug/ml for 1h followed by Alexa Fluor 488 secondary antibody (1 ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom NBP2-81101, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-Fluorescein antibody followed by Alexa Fluor 488 secondary antibody.

Applications for CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free

Application
Recommended Usage

Block/Neutralize

Optimal dilutions of this antibody should be experimentally determined.

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Functional

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This full-length, chimeric rabbit antibody was made using the variable domain sequences of the original Human scFv format, for improved compatibility with existing reagents, assays and techniques. A54 is able to block the CD40-CD154 interaction by around 60-80% and can also induce CD40 sinalling in human B cells when at 'unusually high concentrations' (The CD40 receptor - target, tool and technology, Ellmark, 2002). This scFv can also be used for ELISA studies (PMID: 12153507)

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.

Background: CD40/TNFRSF5

CD40 and its ligand CD154 are members of the tumor necrosis factor receptor (TNFR) and TNF families, respectively, that play key roles in signaling pathways mediating cell growth, survival and differentiation in B-lymphocytes (reviewed in Quezada et al 2004). The CD40 receptor is a 45-50 kDa glycoprotein and is expressed on the surface of B-lymphocytes, some activated T-cells, monocytes, follicular dendritic cells, basal epithelial cells, and in some epithelial and non-epithelial carcinomas. The functions of CD40 have been most extensively studied in B-cells. Ligation of B CD40 by CD154, expressed on activated T cells, stimulates B cell proliferation, differentiation, isotype switching, upregulation of surface molecules contributing to antigen presentation, development of the germinal center, and the humoral memory response. Several distinct structural motifs in the CD40 cytoplasmic domain regaulate various CD40 signaling pathways. A major CD40 signaling pathway activated from CD154 ligand binding is the canonical pathway to the transcription factor family NF-kB, a family of genes mediating immune and inflammatory responses. Although CD40 has been extensively studied as a plasma membrane-associated growth factor membrane receptor. it has also been identified in the cytoplasm and nucleus of normal and neoplastic B-lymphoid cells (Lin-Lee et al. 2006). Other growth factor receptors, including EGF, FGF, and TGF-B have also been identified in the nuclus. It is thought that plasma membrane receptor signaling may be followed by nuclear migration of signaling pathway components. The presence of CD40 in the nucleus of activated normal B lymphocytes and neoplastic B-lymphoid cells suggests that CD40 may play a more complex role in regulating essential growth and survival pathways in B-lymphocytes than previously thought.

Alternate Names

CD40, TNFRSF5

Gene Symbol

CD40

Additional CD40/TNFRSF5 Products

Product Documents for CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for CD40/TNFRSF5 Antibody (A54 scFv) - Chimeric - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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