Product datasheet
Anti-CD3 zeta antibody [EP286Y] ab40804
2 Images
Overview
Product name
Anti-CD3 zeta antibody [EP286Y]
Description
Rabbit monoclonal [EP286Y] to CD3 zeta
Specificity
ab40804 recognises CD3 zeta.
Tested applications
ICC/IF, Flow Cyt
Species reactivity
Reacts with: Human
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within
Human CD3 zeta aa 100 to the C-terminus (C terminal). The exact sequence is proprietary.
Positive control
ICC/IF: Jurkat cells. Flow Cyt: Human peripheral blood lymphocytes.
General notes
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the
following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Alternative versions available:
Anti-CD3 zeta antibody (Alexa Fluor® 488) [EP286Y] (ab197532)
Anti-CD3 zeta antibody (Alexa Fluor® 647) [EP286Y] (ab197037)
Anti-CD3 zeta antibody (HRP) [EP286Y] (ab197038)
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C.
Avoid freeze / thaw cycle.
Storage buffer
PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
Purity
Tissue culture supernatant
Clonality
Monoclonal
Clone number
EP286Y
Isotype
IgG
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Applications
Our Abpromise guarantee covers the use of ab40804 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
ICC/IF
1/250 - 1/500.
Flow Cyt
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with
this antibody.
Application notes
Is unsuitable for WB.
Target
Function
Probable role in assembly and expression of the TCR complex as well as signal transduction
upon antigen triggering.
Involvement in disease
Defects in CD247 are the cause of immunodeficiency due to defect in CD3-zeta (CD3ZID)
[MIM:610163]. An immunological deficiency characterized by T-cells impaired immune response
to alloantigens, tetanus toxoid and mitogens.
Sequence similarities
Belongs to the CD3Z/FCER1G family.
Contains 3 ITAM domains.
Domain
The ITAM domains mediate interaction with SHB.
Post-translational
modifications
Phosphorylated on Tyr residues after T-cell receptor triggering.
Cellular localization
Membrane.
Anti-CD3 zeta antibody [EP286Y] images
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Immunocytochemistry/Immunofluorescence
analysis of Jurkat cells labelling CD3 zeta with
ab40804 at a dilution 1/200. Cells were fixed
with 4% paraformaldehyde and
permeabilized with 0.1% Triton X-100.
ab150077, an Alexa Fluor® 488-conjugated
goat anti-rabbit IgG (1/1000) was used as the
secondary antibody. DAPI (blue) was used as
the nuclear counterstain. ab7291, a mouse
anti-tubulin (1/1000) and ab150120, an Alexa
Fluor® 594-conjugated goat anti-mouse
Immunocytochemistry/ Immunofluorescence -
IgG (1/1000) were used.
Anti-CD3 zeta antibody [EP286Y] (ab40804)
Control 1: primary antibody (1/200) and
secondary antibody, ab150120, an Alexa
Fluor® 594-conjugated goat anti-mouse IgG
(1/1000).
Control 2: ab7291 (1/1000) and secondary
antibody, ab150077, an Alexa Fluor® 488conjugated goat anti-rabbit IgG (1/1000).
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Human peripheral blood lymphocytes stained
with ab40804 (red line). Human whole blood
was processed using a modified protocol
based on Chow et al, 2005 (PMID:
16080188). In brief, human whole blood was
fixed in 4% formaldehyde (methanol-free) for
10 min at 22°C. Red blood cells were then
lyzed by the addition of Triton X-100 (final
concentration - 0.1%) for 15 min at 37°C. For
Flow Cytometry - Anti-CD3 zeta antibody
[EP286Y] (ab40804)
experimentation, cells were treated with 50%
methanol (-20°C) for 15 min at 4°C. Cells
were then incubated with the antibody
(ab40804, 1/1000 dilution) for 30 min at 4°C.
The secondary antibody used was Alexa
Fluor® 488 goat anti-rabbit IgG (H&L)
(ab150077) at 1/2000 dilution for 30 min at
4°C. Isotype control antibody (black line) was
rabbit IgG (monoclonal) (0.1μg/1x106 cells)
used under the same conditions. Unlabelled
sample (blue line) was also used as a control.
Acquisition of >30,000 total events were
collected using a 20mW Argon ion laser
(488nm) and 525/30 bandpass filter. Gating
strategy - peripheral blood lymphocytes.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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