Anti-CD40 [1C10]

Name: Anti-CD40 [1C10]
SKU: Ab02981
Availability: InStock

Description:

Recombinant monoclonal antibody to CD40. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 1C10.

Clone Name:

1C10

Published Application(s):

agonist, IP, ELISA, FC, IF

Published Species Reactivity:

Mouse

Products based on this clone:

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Ab02981-23.0 Anti-CD40 [1C10]
Mouse
Rabbit IgG
Purified
Ships in 4-5 weeks

Ab02981-3.0 Anti-CD40 [1C10]
Mouse
Mouse IgG2b
Purified
Ships in 4-5 weeks

Ab02981-3.3 Anti-CD40 [1C10]
Mouse
Mouse IgG2b
Fc Silent™
Purified
Ships in 4-5 weeks

Ab02981-8.1 Anti-CD40 [1C10]
Mouse
Rat IgG2b
Purified
Ships in 4-5 weeks

Ab02981-8.4 Anti-CD40 [1C10]
Mouse
Rat IgG2b
Fc Silent™
Purified
Ships in 4-5 weeks

Ab02981-1.1 Anti-CD40 [1C10]
Mouse
Mouse IgG1
Purified
In Stock

UniProt Accession Number of Target Protein: P27512

Alternative Name(s) of Target: TNFRSF5; B-cell surface antigen CD40; Bp50; CD40; CD40L receptor; CDW40; MGC9013; T-cell differentiation antigen; Tumor necrosis factor receptor superfamily member 5; tumor necrosis factor receptor superfamily, member 5
Immunogen: The original antibody was generated by immunizing a rat with 20 µg of soluble extracellular domain of recombinant murine CD40 (sCD40).

Specificity: This antibody binds mouse CD40, which is a member of the tumor necrosis factor receptor superfamily. CD40 is an integral membrane protein found on the surface of B lymphocytes.

Application Notes: This antibody was reported to directly induce a specific proliferative response from mature murine B cells, up-regulate several B cell surface antigens, and rescue immature B lymphoma cells from anti-IgM-induced growth arrest. This initial screening of hybridoma supernatants for reactivity towards CD40 transfected L cells was done using flow cytometry. This antibody recognized recombinant CD40 expressed on L cells and the soluble extracellular domain of murine CD40 coated onto microtiter plates in an ELISA. This antibody was also used in the immunofluorescent staining of adult murine spleen cells. The staining was restricted to the B220+ population. This antibody also specifically immunoprecipitated a 45-kDa protein from the surface of purified murine splenic B cells (PMID: 7519997). This antibody was capable of blocking binding of FGK45 to CD40 on murine B cells in vitro (PMID: 24416732). The mouse IgG1 version of this antibody had a tenfold higher affinity for mouse FcγRIIB as compared to mouse IgG2a or the parental rat IgG2b (PMID: 21852502). The mouse IgG1 version of this antibody was used in the treatment of M38 tumor bearing mice and it was reported that there was a strong increase in PD-L1 expression both on tumor-infiltrating monocytes and TAMs (PMID: 25623164).

Antibody first published in: Heath et al. Monoclonal antibodies to murine CD40 define two distinct functional epitopes. Eur J Immunol. 1994 Aug;24(8):1828-34. PMID:7519997
Note on publication: This paper discusses the generation and functional characterization of two antibodies that recognize two distinct CD40 epitopes.

Western blot using Ab02981-1.1 Click on image for more details.

Western Blot using Anti-CD40 antibody 1C10 (Ab02981). Mouse spleen(A) (2µg/ml), thymus(B) (2µg/ml), mouse lymph node(C) (1ug/ml) and lung(D) (1ug/ml) tissue lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric mouse version of 1C10 (Ab02981-1.1) before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

IF staining of mouse splenocytes cells using Ab02981-1.1. Click on image for more details.

Immunofluorescence staining of mouse splenocytes cells with Anti-CD40 (Ab02981) 1C10 Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes cells immobilized on Shi-fix™ plus cover-slips stained with the chimeric mouse IgG1 version of 1C10 (Ab02981-1.1) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining in subset of cells. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab02981-1.1, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-1.1) followed by staining with Alexa Fluor® 488 secondary antibody.

Flow cytometry analysis of mouse splenocytes using Ab02981-1.1. Click on image for more details.

Flow cytometry using the Anti-CD40 antibody 1C10 (Ab02981). Paraformaldehyde fixed mouse splenocytes were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of 1C10 (Ab02981-1.1, 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.

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