United Kingdom (UK)
Recombinant monoclonal antibody to CD3 epsilon. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma YTH 12.5.
UniProt Accession Number of Target Protein: P07766
Alternative Name(s) of Target: CD3e; T-cell surface glycoprotein CD3 epsilon chain; T-cell surface antigen T3/Leu-4 epsilon chain; CD3e; CD3E; T3E; YTH12.5
Immunogen: Human CD3.
Specificity: Recognises the epsilon chain of the human CD3 antigen complex on human T cells.
Antibody first published in:
Clark M, Hale G, Waldmann H. Interaction of rat monoclonal antibodies with human killer cells. Adv Exp Med Biol. 1985;186:797-803. PMID:4050599
Note on publication:
Describes the production of rat monoclonal antibody and the assessment of its potential to mediate immune effector mechanisms.
Flow-cytometry using the anti-CD3E antibody YTH 12.5 (Ab00170) Jurkat cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of YTH 12.5 (Ab00170-23.0, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.
Immunohistochemical staining of human tonsil tissue using anti-CD3E antibody (Ab00170) YTH 12.5 Anti-CD3E staining of paraffin embedded human tissue using the rabbit-chimeric version of YTH 12.5 (Ab00170-23.0). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer (Dako, cat. #X0909). Primary antibody incubation with Ab00170-23.0 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody (Dako cat#K4009) for 20 mins followed by DAB (3,3′-diaminobenzidine), and counter-staining with haemotoxylin. Strong membrane staining of T-cells at the periphery of the germinal centre and between follicles may be observed. Recommended concentration, 2-4 µg/ml.
Flow-cytometry using anti-CD3 epsilon antibody YTH 12.5 (Ab00170) Human lymphocytes were stained with an isotype control (Ab00102-23.0, panel A) or the rabbit-chimeric version of YTH 12.5 (Ab00170-23.0, panel B) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using a AF488 conjugated donkey anti-rabbit antibody (ab150073) and cells analysed on a FACSCanto flow-cytometer.
Flow-cytometry using anti-CD3 antibody YTH 12.5 (Ab00170) Rhesus monkey lymphocytes were stained with an isotype control (Ab00102-23.0, red) or the rabbit-chimeric version of YTH 12.5 (Ab00170-23.0, blue) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using a AF488 conjugated donkey anti-rabbit antibody (ab150073) and cells analysed on a FlowJo single-cell flow cytometer.
Western Blot using anti-CD3E antibody YTH 12.5 (Ab00170) Human spleen sample (35µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of YTH 12.5 (Ab00170-23.0) at 0.3 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for CD3E is 23.1 kDa. Ab00170-23.0 successfully detected human CD3E.
Flow-cytometry using the anti-CD3E antibody YTH 12.5 (Ab00170) Jurkat cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of YTH 12.5 (Ab00170-23.0, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.
Immunohistochemical staining of human tonsil tissue using anti-CD3E antibody (Ab00170) YTH 12.5 Anti-CD3E staining of paraffin embedded human tissue using the rabbit-chimeric version of YTH 12.5 (Ab00170-23.0). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer (Dako, cat. #X0909). Primary antibody incubation with Ab00170-23.0 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody (Dako cat#K4009) for 20 mins followed by DAB (3,3′-diaminobenzidine), and counter-staining with haemotoxylin. Strong membrane staining of T-cells at the periphery of the germinal centre and between follicles may be observed. Recommended concentration, 2-4 µg/ml.