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- Ab00914-2.0 Anti-CD3e [898H2-6-15]
- Pig
- Mouse IgG2a
- Purified
- In Stock
- Ab00914-2.3 Anti-CD3e [898H2-6-15]
- Pig
- Mouse IgG2a
- Fc Silent™
- Purified
- Ships in 4-5 weeks
- Ab00914-23.0 Anti-CD3e [898H2-6-15]
- Pig
- Rabbit IgG
- Purified
- In Stock
Recombinant monoclonal antibody to CD3e. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 898H2-6-15.
UniProt Accession Number of Target Protein: Q7YRN2
Alternative Name(s) of Target: Leu4/T3; Cluster of Differentiation 3 epsilon chain; T cell glycoprotein CD3 epsilon chain
Immunogen: Murine antibodies were produced by immunising C3H/HeJ mice with swine peripheral blood lymphocytes. Hybridomas were generated by obtaining splenocytes from the immunised mice and fusing these with the non-Ig-producing myeloma cell line P3X63Ag8.653 via established methods.
Specificity: Hybridoma 898H2-6-15 was shown via double-colour flow cytometry analysis to bind swine lymphocyte populations specifically. It was further shown using FACS that purified 898H2-6-15 bound an epitope on CD3e shared by another anti-pig CD3 Ab (STH164). CD3, and the e chain in particular, is crucial for T cell receptor-dependent signal transduction across the lymphocyte membrane.
Application Notes: The initial identification of 898H2-6-15 showed that it recognised the T-cell receptor-associated glycoprotein CD3, in particular the e chain (Huang et al, 1999). More recently, the 898H2-6-15 VH and VL domains were reformatted into a scFv and conjugated to diphtheria toxin (Wang et al, 2011). This version of the antibody as an 'immunotoxin' has been shown to deplete circulating T cells in swine without observation of any clinical toxicities.
Antibody first published in:
Huang et al. Characterization of a monoclonal anti-porcine CD3 antibody Xenotransplantation 1999: 5: 201-212 PMID:10503787
Note on publication:
Describes the identification and qualitative characterisation of murine anti-pig CD3e antibodies from hybridoma. These were shown to be specific for pig T lymphocytes, as opposed to human and baboon peripheral blood cells, and mouse spleen cells.
Immunofluorescence staining of pig peripheral blood monocytes with anti-CD3e (Ab00914) 898H2-6-15 Immunofluorescence analysis of paraformaldehyde fixed pig peripheral blood monocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 898H2-6-15 (Ab00914-23.0) 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 Ab00914-23.0, 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 using Anti-CD3e antibody 898H2-6-15 (Ab00914). Pig spleen(A) (1µg/ml), pig thymus(B) (0.1µg/ml) and lymph node(C) (0.1µg/ml) tissue lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of 898H2-6-15 (Ab00914-23.0) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow cytometry using the Anti-CD3e antibody 898H2-6-15 (Ab00914). Paraformaldehyde fixed pig peripheral blood monocytes permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of 898H2-6-15 (Ab00914-23.0, 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.
Immunofluorescence staining of pig peripheral blood monocytes with anti-CD3e (Ab00914) 898H2-6-15 Immunofluorescence analysis of paraformaldehyde fixed pig peripheral blood monocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 898H2-6-15 (Ab00914-23.0) 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 Ab00914-23.0, 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.