Anti-CD226 [10E5]

Description:

Recombinant monoclonal antibody to CD226. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 10E5.

Clone Name:

10E5

Published Application(s):

Published Species Reactivity:

Mouse

Products based on this clone:

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Ab01366-8.1 Anti-CD226 [10E5]
Mouse
Rat IgG2b
Purified
In Stock

Ab01366-8.4 Anti-CD226 [10E5]
Mouse
Rat IgG2b
Fc Silent™
Purified
Ships in 4-5 weeks

Ab01366-1.1 Anti-CD226 [10E5]
Mouse
Mouse IgG1
Purified
Ships in 4-5 weeks

Ab01366-2.0 Anti-CD226 [10E5]
Mouse
Mouse IgG2a
Purified
Ships in 4-5 weeks

Ab01366-2.3 Anti-CD226 [10E5]
Mouse
Mouse IgG2a
Fc Silent™
Purified, VivopureX
In Stock

Ab01366-23.0 Anti-CD226 [10E5]
Mouse
Rabbit IgG
Purified
In Stock

UniProt Accession Number of Target Protein: Q8K4F0

Alternative Name(s) of Target: DNAM-1; PTA1; PTA-1; platelet and T cell activation antigen 1; TLISA1
Immunogen: This antibody was raised by immunising Lewis and Lou/M rats with the murine Th1 cell clone AE7 and Th1 cell lines derived from 5B6 and DO11.10 TCR transgenic mice, before screening for reactivity with Th1 clones and a lack of reactivity to Th2/Th0 cell lines.

Specificity: This antibody is specific for murine CD226.

Application Notes: The specificity of this antibody for murine CD226 has been confirmed in flow cytometric analysis using 293 cells transfected with CD226 cDNA and untransfected 293 cells (Dardalhon et al, 2005). This antibody has been used in flow cytometry to identify the surface expression pattern of CD226, which is predominantly confined to T cells (Dardalhon et al, 2005; Yasuma et al, 2016), and to detect NK cell CD226 expression and apoptosis (Huang et al, 2011). Immunisation of mice with this antibody inhibits antigen-specific T cell expansion (Dardalhon et al, 2005). Additionally, treatment of experimental autoimmune encephalomyelitis (EAE) mice with this antibody delays the onset and reduces the severity of Th1 cell-mediated autoimmune disease (Dardalhon et al, 2005).

Antibody first published in: Dardalhon et al. CD226 is specifically expressed on the surface of Th1 cells and regulates their expansion and effector functions. J Immunol. 2005 Aug 1;175(3):1558-65. PMID:16034094
Note on publication: Describes the original generation of this antibody, and its characterisation.

Western blot on mouse spleen tissue lysate. Using Ab01366-23.0. Click on image for more details.

Western Blot using anti-CD266 antibody 10E5 (Ab01366) Mouse spleen tissue lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of 10E5 (Ab01366-23.0) at 2 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

IF staining of mouse splenocytes using Ab01366-23.0. Click on image for more details.

Immunofluorescence staining of fixed mouse splenocytes with anti- CD226 antibody 10E5 (Ab01366) Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 10E5 (Ab01366-23.0) 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 Ab01366-23.0, 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 Ab01366-23.0. Click on image for more details.

Flow cytometry using the Anti-CD226 antibody 10E5 (Ab01366). Paraformaldehyde fixed mouse splenocytes were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of 10E5 (Ab01366-23.0, 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.

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