Anti-CD40 [FGK45]

Name: Anti-CD40 [FGK45]
SKU: Ab01627
Availability: InStock

Description:

Recombinant monoclonal antibody to CD40. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma FGK45.

Clone Name:

FGK45

Published Application(s):

Activating, agonist, functional assays, WB, FC

Published Species Reactivity:

Mouse

Products based on this clone:

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Ab01627-7.1 Anti-CD40 [FGK45]
Mouse
Rat IgG2a
Purified
In Stock

Ab01627-7.4 Anti-CD40 [FGK45]
Mouse
Rat IgG2a
Fc Silent™
Purified
In Stock

Ab01627-3.0 Anti-CD40 [FGK45]
Mouse
Mouse IgG2b
Purified
Ships in 4-5 weeks

Ab01627-3.3 Anti-CD40 [FGK45]
Mouse
Mouse IgG2b
Fc Silent™
Purified
Ships in 4-5 weeks

Ab01627-23.0 Anti-CD40 [FGK45]
Mouse
Rabbit IgG
Purified
In Stock

Ab01627-1.1 Anti-CD40 [FGK45]
Mouse
Mouse IgG1
Purified
In Stock

UniProt Accession Number of Target Protein: P27512

Alternative Name(s) of Target: Tnfrsf5; Tumor necrosis factor receptor superfamily member 5; B-cell surface antigen CD40; Bp50; CD40L receptor; FGK4.5
Immunogen: The original rat IgG2a antibody was raised against the recombinant mouse CD40 fusion protein.

Specificity: This antibody recognizes CD40. It is a type I transmembrane glycoprotein that belongs to the tumor necrosis factor receptor (TNFR) superfamily. CD40 is expressed broadly on antigen-presenting cells (APCs) such as dendritic cells, B cells, macrophages, and monocytes as well as non-immune endothelial cells, basal epithelial cells, and a range of tumors. Upon binding to its ligand CD154, CD40 acts as a costimulatory molecule for the activation of B cells, dendritic cells, monocytes, and other APCs.

Application Notes: FGK45 is an agonistic antibody that has been shown to activate CD40 expressing antigen presenting cells (APCs) what might promote anti-tumor T cell responses. For instance, the rat IgG2a version of this antibody was used to promote maturation of APCs what improved the ability of the macrophages to fight a tumour (Luheshi et al., 2016; pmid: 26918344). Another group used the same antibody to stimulate T-cells in a study on a tumor vaccine (Bartkowiak et al., 2015; pmid: 26351680). Others used ths clone as well to induce potent anti-tumor activity (Carmi et al., 2015; pmid: 25924063). On the other hand, it can also be used to inhibit CD40/CD154 interaction in vitro and in vivo. Furthermore, this clone can be also utilized to identify CD40-positive cells by flow cytometry (Rolink et al., 1996; pmid: 8885865). Administering this antibody can be also utilized to induce inflamatory bowel disease-like characteristics and create a mouse IBD model to study this class of diseases (Bauche et al., 2018; pmid: 30097293).

Antibody first published in: Rolink et al. The SCID but not the RAG-2 gene product is required for S mu-S epsilon heavy chain class switching Immunity. 1996 Oct;5(4):319-30. doi: 10.1016/s1074-7613(00)80258-7. PMID:8885865
Note on publication: Describes the generation and characterization of the antibody FGK45.

IF staining of mouse splenocytes using Ab01627-23.0. Click on image for more details.

Immunofluorescence staining of mouse splenocytes with anti-CD40 (Ab01627) FGK45 Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of FGK45 (Ab01627-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01627-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.

Western blot on mouse lymph node tissue lysate using Ab01627-23.0. Click on image for more details.

Western Blot using Anti-CD40 antibody FGK45 (Ab01627). Mouse lymph node tissue lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of FGK45 (Ab01627-23.0) at 2µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

Flow cytometry analysis of mouse splenocytes using Ab01627-23.0. Click on image for more details.

Flow cytometry using the Anti-CD40 antibody FGK45 (Ab01627). Paraformaldehyde fixed mouse splenocytes were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of FGK45 (Ab01627-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

IF staining of mouse splenocytes using Ab01627-23.0. Click on image for more details.

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