Product: CD3 zeta Antibody
Catalog: AF5405
Description: Rabbit polyclonal antibody to CD3 zeta
Application: WB IF/ICC
Reactivity: Human, Mouse
Prediction: Pig, Bovine, Horse, Sheep, Rabbit
Mol.Wt.: 19 kDa; 19kD(Calculated).
Uniprot: P20963
RRID: AB_2837889

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse
Prediction:
Pig(100%), Bovine(100%), Horse(90%), Sheep(90%), Rabbit(100%)
Clonality:
Polyclonal
Specificity:
CD3 zeta Antibody detects endogenous levels of total CD3 zeta.
RRID:
AB_2837889
Cite Format: Affinity Biosciences Cat# AF5405, RRID:AB_2837889.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

4930549J05Rik; A430104F18Rik; AW552088; CD247; CD247 antigen; Cd247 molecule; Cd3; CD3 antigen, zeta polypeptide, isoform CRA_b; CD3 antigen, zeta subunit; CD3-eta; CD3H; CD3Q; Cd3z; CD3Z antigen zeta polypeptide (TiT3 complex); CD3Z_HUMAN; CD3zeta; CD3zeta chain; MGC140430; T cell receptor T3 zeta chain; T cell surface glycoprotein CD3 zeta chain; T-cell antigen receptor complex, zeta subunit of CD3; T-cell receptor T3 zeta chain; T-cell surface glycoprotein CD3 zeta chain; T3z; TCR zeta chain; TCRk; TCRZ; TCRzeta;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P20963 CD3Z_HUMAN:

CD3Z is expressed in normal lymphoid tissue and in peripheral blood mononuclear cells (PBMCs) (PubMed:11722641).

Sequence:
MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Rabbit
100
Horse
90
Sheep
90
Zebrafish
78
Xenopus
75
Chicken
70
Dog
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P20963 As Substrate

Site PTM Type Enzyme
S58 Phosphorylation
Y64 Phosphorylation
Y72 Phosphorylation
Y83 Phosphorylation
K88 Ubiquitination
K99 Ubiquitination
K104 Ubiquitination
Y111 Phosphorylation P06241 (FYN)
K116 Ubiquitination
K118 Ubiquitination
Y123 Phosphorylation P06241 (FYN)
S124 Phosphorylation
K129 Ubiquitination
K136 Ubiquitination
Y142 Phosphorylation P06239 (LCK)
S146 Phosphorylation
T147 Phosphorylation
T149 Phosphorylation
K150 Ubiquitination
T152 Phosphorylation
Y153 Phosphorylation

Research Backgrounds

Function:

Part of the TCR-CD3 complex present on T-lymphocyte cell surface that plays an essential role in adaptive immune response. When antigen presenting cells (APCs) activate T-cell receptor (TCR), TCR-mediated signals are transmitted across the cell membrane by the CD3 chains CD3D, CD3E, CD3G and CD3Z. All CD3 chains contain immunoreceptor tyrosine-based activation motifs (ITAMs) in their cytoplasmic domain. Upon TCR engagement, these motifs become phosphorylated by Src family protein tyrosine kinases LCK and FYN, resulting in the activation of downstream signaling pathways. CD3Z ITAMs phosphorylation creates multiple docking sites for the protein kinase ZAP70 leading to ZAP70 phosphorylation and its conversion into a catalytically active enzyme. Plays an important role in intrathymic T-cell differentiation. Additionally, participates in the activity-dependent synapse formation of retinal ganglion cells (RGCs) in both the retina and dorsal lateral geniculate nucleus (dLGN) (By similarity).

PTMs:

Phosphorylated on Tyr residues after T-cell receptor triggering by LCK in association with CD4/CD8.

Subcellular Location:

Membrane>Single-pass type I membrane protein.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

CD3Z is expressed in normal lymphoid tissue and in peripheral blood mononuclear cells (PBMCs).

Subunit Structure:

The TCR-CD3 complex is composed of a CD3D/CD3E and a CD3G/CD3E heterodimers that preferentially associate with TCRalpha and TCRbeta, respectively, to form TCRalpha/CD3E/CD3G and TCRbeta/CD3G/CD3E trimers. In turn, the hexamer interacts with CD3Z homodimer to form the TCR-CD3 complex. Alternatively, TCRalpha and TCRbeta can be replaced by TCRgamma and TCRdelta. Interacts with SLA. Interacts with TRAT1. Interacts with DOCK2. Interacts with SLA2. Interacts with SHB. Interacts with ZAP70. Interacts (tyrosine phosphorylated) with SHC1 (via SH2 domain). Interacts with PTPRC. Interacts with CRK; this interaction regulates CD3Z phosphorylation. Interacts (on T cell side) with CD81, ICAM1 and CD9 at immunological synapses between antigen-presenting cells and T cells. Interacts with CD160.

(Microbial infection) Interacts with HIV-1 Nef; this interaction up-regulates the expression of the Fas ligand (FASLG) at the cell surface.

(Microbial infection) Interacts with HIV-2 Nef protein; this interaction induces down-regulation of cell surface TCR/CD3 complexes.

Family&Domains:

The ITAM domains mediate interaction with SHB.

Belongs to the CD3Z/FCER1G family.

Research Fields

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.   (View pathway)

· Organismal Systems > Immune system > Th1 and Th2 cell differentiation.   (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation.   (View pathway)

· Organismal Systems > Immune system > T cell receptor signaling pathway.   (View pathway)

References

1). Lupeol ameliorates LPS/D-GalN induced acute hepatic damage by suppressing inflammation and oxidative stress through TGFβ1-Nrf2 signal pathway. Aging-US, 2021 (PubMed: 33707345) [IF=5.2]

Application: IF/ICC    Species: Mice    Sample: liver tissue

Figure 3 Lupeol restrains LPS/GalN-induced liver inflammation. (A) Immunohistochemistry staining of VCAM-1 in zebrafish, n=20, figures are magnified as 400x, bar=200μm. (B) Quantitative analysis of VCAM-1, data are represented as mean ± SEM. (C) Expression of TNF-α and VCAM-1 in mice. (D) Quantitative analysis of TNF-α and VCAM-1 protein. (E, F) Immunohistochemistry analysis for TNF-α location and expression in mice. (G, H) Immunofluorescence analysis for CD3 expression in mice. (I, J) Immunohistochemistry staining of F4/80 and LY-6G in mice and quantification of F4/80 and LY-6G expression. Data are shown as the mean ± SEM, n=3-4 group, *P<0.05, **P<0.01, ***P<0.001, control group vs other groups. #P<0.05, ##P<0.01, ### P<0.001. Figures are magnified as 200x, bar=50μm.

2). The Third Generation Anti-HER2 Chimeric Antigen Receptor Mouse T Cells Alone or Together With Anti-PD1 Antibody Inhibits the Growth of Mouse Breast Tumor Cells Expressing HER2 in vitro and in Immune Competent Mice. Frontiers in Oncology, 2020 (PubMed: 32766150) [IF=4.7]

Application: WB    Species: mouse    Sample: CAR-T cells

FIGURE 2 | Preparation of the third generation anti-HER2 CAR-T cells. (D) The expression level of CAR (containing exogenous CD3ζ) in CAR-T cells was detected by western blotting using an anti-CD3ζ antibody, with blank T cells as negative control, and endogenous CD3ζ as a loading control. The experiments have been repeated for three times.CAR, chimeric antigen receptor; scFv, single chain variable fragment; VH, heavy chain variable region; VL, light chain variable region; MOI, multiplicity of infection;7-AAD, 7-aminoactinomycin D; Blank T, T cells not transduced; CAR-T, T cells transduced with CAR.

3). Interleukin 35 induced Th2 and Tregs bias under normal conditions in mice. PeerJ, 2018 (PubMed: 30258726) [IF=2.7]

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Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

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