Anti-CD3 epsilon [YTH 12.5]

SKU: orb411552

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Description

Rat monoclonal antibody to CD3 epsilon

Images & Validation

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Item 1 of 5

Tested Applications	IF, IHC, WB
Reactivity	Human

Key Properties

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Host	Rat
Clonality	Monoclonal
Isotype	Rat IgG
Clone No.	YTH 12.5
Immunogen	Human CD3.
Target	CD3 epsilon
Purity	Purified
Conjugation	Unconjugated

Storage & Handling

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Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Buffer/Preservatives	PBS with 0.02% Proclin 300.
Concentration	1 mg/ml
Disclaimer	For research use only

Alternative Names

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CD3e; T-cell surface glycoprotein CD3 epsilon chain; T-cell surface antigen T3/Leu-4 epsilon chain; CD3e; CD3E; T3E; YTH12.5

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Flow-cytometry using anti-CD3 antibody YTH 12.5 Rhesus monkey lymphocytes were stained with an isotype control (orb256390, red) or the rabbit-chimeric version of YTH 12.5 (orb411552, 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 and cells analysed on a FlowJo single-cell flow cytometer.

Flow-cytometry using anti-CD3 epsilon antibody YTH 12.5 Human lymphocytes were stained with an isotype control (orb256390, panel A) or the rabbit-chimeric version of YTH 12.5 (orb411552, 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 and cells analysed on a FACSCanto flow-cytometer.

Flow-cytometry using the anti-CD3E antibody YTH 12.5 Jurkat cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of YTH 12.5 (orb411552, 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 YTH 12.5 Anti-CD3E staining of paraffin embedded human tissue using the rabbit-chimeric version of YTH 12.5 (orb411552). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer. Primary antibody incubation with orb411552 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody 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.

Western Blot using anti-CD3E antibody YTH 12.5 Human spleen sample (35 µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of YTH 12.5 (orb411552) 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. orb411552 successfully detected human CD3E.

Quick Database Links

Gene Symbol

CD3 epsilon

UniProt

P07766

UniProt Details

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No UniProt data available

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Protocol Information

Western Blot (IB, immunoblot)

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IHC

Immunohistochemistry

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Immunofluorescence

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Find More Protocols

Routledge, E. G. et al. A humanized monovalent CD3 antibody which can activate homologous complement Eur J Immunol, 21, 2717-2725 (1991)

PubMed: 1834468