CD3 zeta Antibody | Affinity Biosciences

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FIGURE 2 | Preparation of the third generation anti-HER2 CAR-T cells.

Figure 3 Lupeol restrains LPS/GalN-induced liver inflammation.

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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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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

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Related Downloads

MS Report

HPLC Report

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

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:

P20963(CD3Z_HUMAN) >>Visit HPA database.

Gene(ID):

CD247(919)

Expression:

P20963 CD3Z_HUMAN:

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

Sequence:

P20963 CD3Z_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

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

Sheep

Zebrafish

Xenopus

Chicken

Dog

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P20963 As Substrate

P20963

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

Research Backgrounds

P20963_CD3Z_HUMAN

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.

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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CD3 zeta Antibody - #AF5405