Product: ZAP70 Antibody
Catalog: AF6312
Description: Rabbit polyclonal antibody to ZAP70
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 70kDa; 70kD(Calculated).
Uniprot: P43403
RRID: AB_2835171

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, 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,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
ZAP70 Antibody detects endogenous levels of total ZAP70.
RRID:
AB_2835171
Cite Format: Affinity Biosciences Cat# AF6312, RRID:AB_2835171.
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

70 kDa zeta associated protein; 70 kDa zeta-associated protein; EC 2.7.10.2; FLJ17670; FLJ17679; Selective T cell defect; SRK; STD; Syk related tyrosine kinase; Syk-related tyrosine kinase; Truncated ZAP kinase; Tyrosine protein kinase ZAP70; Tyrosine-protein kinase ZAP-70; TZK; ZAP 70; ZAP70; ZAP70_HUMAN; Zeta chain associated protein kinase 70kD; Zeta chain associated protein kinase 70kDa; Zeta chain associated protein kinase 70kDa isoform 1; Zeta chain associated protein kinase 70kDa isoform 2; Zeta chain TCR associated protein kinase 70kD; Zeta chain TCR associated protein kinase 70kDa;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P43403 ZAP70_HUMAN:

Expressed in T- and natural killer cells. Also present in early thymocytes and pro/pre B-cells.

Description:
ZAP70 (human) a tyrosine kinase of the Syk family. Associates with the T-cell antigen receptor zeta-chain after TCR stimulation. Phosphorylated by Src-family kinases following antigen receptor activation. Plays a role in lymphocyte activation.
Sequence:
MPDPAAHLPFFYGSISRAEAEEHLKLAGMADGLFLLRQCLRSLGGYVLSLVHDVRFHHFPIERQLNGTYAIAGGKAHCGPAELCEFYSRDPDGLPCNLRKPCNRPSGLEPQPGVFDCLRDAMVRDYVRQTWKLEGEALEQAIISQAPQVEKLIATTAHERMPWYHSSLTREEAERKLYSGAQTDGKFLLRPRKEQGTYALSLIYGKTVYHYLISQDKAGKYCIPEGTKFDTLWQLVEYLKLKADGLIYCLKEACPNSSASNASGAAAPTLPAHPSTLTHPQRRIDTLNSDGYTPEPARITSPDKPRPMPMDTSVYESPYSDPEELKDKKLFLKRDNLLIADIELGCGNFGSVRQGVYRMRKKQIDVAIKVLKQGTEKADTEEMMREAQIMHQLDNPYIVRLIGVCQAEALMLVMEMAGGGPLHKFLVGKREEIPVSNVAELLHQVSMGMKYLEEKNFVHRDLAARNVLLVNRHYAKISDFGLSKALGADDSYYTARSAGKWPLKWYAPECINFRKFSSRSDVWSYGVTMWEALSYGQKPYKKMKGPEVMAFIEQGKRMECPPECPPELYALMSDCWIYKWEDRPDFLTVEQRMRACYYSLASKVEGPPGSTQKAEAACA

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
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Chicken
100
Rabbit
100
Zebrafish
63
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P43403 As Substrate

Site PTM Type Enzyme
Y12 Phosphorylation
K25 Ubiquitination
Y46 Phosphorylation
Y69 Phosphorylation P06239 (LCK)
K75 Ubiquitination
Y87 Phosphorylation
K100 Ubiquitination
S106 Phosphorylation
Y126 Phosphorylation P43403 (ZAP70) , P06239 (LCK)
K132 Ubiquitination
K151 Ubiquitination
Y164 Phosphorylation
S166 Phosphorylation
S167 Phosphorylation
K176 Ubiquitination
Y178 Phosphorylation P06239 (LCK)
S179 Phosphorylation
K186 Ubiquitination
K193 Ubiquitination
T197 Phosphorylation
Y198 Phosphorylation
Y204 Phosphorylation
K206 Ubiquitination
Y209 Phosphorylation
Y211 Phosphorylation
K217 Ubiquitination
K220 Ubiquitination
Y221 Phosphorylation
K242 Ubiquitination
Y248 Phosphorylation
K251 Ubiquitination
S257 Phosphorylation
S258 Phosphorylation
S263 Phosphorylation
T278 Phosphorylation
T286 Phosphorylation
S289 Phosphorylation
Y292 Phosphorylation P06239 (LCK) , P43403 (ZAP70)
T293 Phosphorylation
T300 Phosphorylation
S301 Phosphorylation
K304 Ubiquitination
T312 Phosphorylation
S313 Phosphorylation
Y315 Phosphorylation P06239 (LCK) , P43403 (ZAP70)
S317 Phosphorylation
Y319 Phosphorylation P00519 (ABL1) , P43403 (ZAP70) , P06239 (LCK)
S320 Phosphorylation
K326 Ubiquitination
K329 Ubiquitination
S351 Phosphorylation
K361 Ubiquitination
K362 Ubiquitination
K369 Ubiquitination
K372 Ubiquitination
K377 Ubiquitination
Y397 Phosphorylation
K450 Ubiquitination
Y451 Phosphorylation
K455 Ubiquitination
Y474 Phosphorylation P06239 (LCK)
K476 Ubiquitination
K484 Ubiquitination
S491 Phosphorylation
Y492 Phosphorylation P06239 (LCK) , P43403 (ZAP70)
Y493 Phosphorylation P06239 (LCK) , P43403 (ZAP70)
T494 Phosphorylation
S497 Phosphorylation
K500 Ubiquitination
K504 Ubiquitination
Y506 Phosphorylation
S520 Phosphorylation
S534 Phosphorylation
Y535 Phosphorylation
K538 Ubiquitination
K544 Ubiquitination
K556 Ubiquitination
Y569 Phosphorylation
Y597 Phosphorylation
Y598 Phosphorylation
S599 Phosphorylation
K603 Acetylation
K603 Ubiquitination
T611 Phosphorylation
K613 Ubiquitination

PTMs - P43403 As Enzyme

Substrate Site Source
O43561-4 (LAT) Y126 Uniprot
O43561-2 (LAT) Y127 Uniprot
O43561-4 (LAT) Y131 Uniprot
O43561-2 (LAT) Y132 Uniprot
O43561-1 (LAT) Y156 Uniprot
O43561 (LAT) Y161 Uniprot
O43561-3 (LAT) Y163 Uniprot
O43561-3 (LAT) Y168 Uniprot
O43561-4 (LAT) Y170 Uniprot
O43561-2 (LAT) Y171 Uniprot
O43561-4 (LAT) Y190 Uniprot
O43561-2 (LAT) Y191 Uniprot
O43561-1 (LAT) Y200 Uniprot
O43561-3 (LAT) Y207 Uniprot
O43561-1 (LAT) Y220 Uniprot
O43561-4 (LAT) Y225 Uniprot
O43561-2 (LAT) Y226 Uniprot
O43561-3 (LAT) Y227 Uniprot
O43561 (LAT) Y255 Uniprot
O43561-3 (LAT) Y262 Uniprot
P15941-7 (MUC1) Y229 Uniprot
P15941-11 (MUC1) Y238 Uniprot
P15941 (MUC1) Y1203 Uniprot
P19174 (PLCG1) Y783 Uniprot
P19174 (PLCG1) Y1253 Uniprot
P29353-7 (SHC1) Y239 Uniprot
P29353-2 (SHC1) Y240 Uniprot
P29353-2 (SHC1) Y317 Uniprot
P29353 (SHC1) Y349 Uniprot
P29353 (SHC1) Y350 Uniprot
P29353 (SHC1) Y427 Uniprot
P43403-2 (ZAP70) Y8 Uniprot
P43403-2 (ZAP70) Y12 Uniprot
P43403 (ZAP70) Y126 Uniprot
P43403 (ZAP70) Y292 Uniprot
P43403 (ZAP70) Y315 Uniprot
P43403 (ZAP70) Y319 Uniprot
P43403 (ZAP70) Y492 Uniprot
P43403 (ZAP70) Y493 Uniprot
P51452 (DUSP3) Y38 Uniprot
P51452 (DUSP3) Y138 Uniprot
Q13094 (LCP2) Y113 Uniprot
Q13094 (LCP2) Y128 Uniprot
Q13094 (LCP2) Y145 Uniprot
Q13094 (LCP2) Y423 Uniprot
Q13094 (LCP2) Y426 Uniprot
Q16539 (MAPK14) Y323 Uniprot
Q8N1K5 (THEMIS) Y540 Uniprot
Q8N1K5 (THEMIS) Y541 Uniprot
Q9UJU6 (DBNL) Y334 Uniprot
Q9UJU6-2 (DBNL) Y335 Uniprot
Q9UJU6 (DBNL) Y344 Uniprot
Q9UJU6-2 (DBNL) Y345 Uniprot
Q9UQC2 (GAB2) Y614 Uniprot
Q9UQQ2 (SH2B3) Y273 Uniprot

Research Backgrounds

Function:

Tyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates motility, adhesion and cytokine expression of mature T-cells, as well as thymocyte development. Contributes also to the development and activation of primary B-lymphocytes. When antigen presenting cells (APC) activate T-cell receptor (TCR), a serie of phosphorylations lead to the recruitment of ZAP70 to the doubly phosphorylated TCR component CD247/CD3Z through ITAM motif at the plasma membrane. This recruitment serves to localization to the stimulated TCR and to relieve its autoinhibited conformation. Release of ZAP70 active conformation is further stabilized by phosphorylation mediated by LCK. Subsequently, ZAP70 phosphorylates at least 2 essential adapter proteins: LAT and LCP2. In turn, a large number of signaling molecules are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Furthermore, ZAP70 controls cytoskeleton modifications, adhesion and mobility of T-lymphocytes, thus ensuring correct delivery of effectors to the APC. ZAP70 is also required for TCR-CD247/CD3Z internalization and degradation through interaction with the E3 ubiquitin-protein ligase CBL and adapter proteins SLA and SLA2. Thus, ZAP70 regulates both T-cell activation switch on and switch off by modulating TCR expression at the T-cell surface. During thymocyte development, ZAP70 promotes survival and cell-cycle progression of developing thymocytes before positive selection (when cells are still CD4/CD8 double negative). Additionally, ZAP70-dependent signaling pathway may also contribute to primary B-cells formation and activation through B-cell receptor (BCR).

PTMs:

Phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Phosphorylation of Tyr-315 and Tyr-319 are essential for ZAP70 positive function on T-lymphocyte activation whereas Tyr-292 has a negative regulatory role. Within the C-terminal kinase domain, Tyr-492 and Tyr-493 are phosphorylated after TCR induction, Tyr-492 playing a negative regulatory role and Tyr-493 a positive. Tyr-493 is dephosphorylated by PTN22.

Ubiquitinated in response to T cell activation. Deubiquitinated by OTUD7B.

Subcellular Location:

Cytoplasm. Cell membrane>Peripheral membrane protein.
Note: In quiescent T-lymphocytes, it is cytoplasmic. Upon TCR activation, it is recruited at the plasma membrane by interacting with CD247/CD3Z. Colocalizes together with RHOH in the immunological synapse. RHOH is required for its proper localization to the cell membrane and cytoskeleton fractions in the thymocytes (By similarity).

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

Expressed in T- and natural killer cells. Also present in early thymocytes and pro/pre B-cells.

Subunit Structure:

Interacts with CD247/CD3Z; this interaction docks ZAP70 at the stimulated TCR. Interacts with NFAM1. Interacts with adapter protein SLA; this interaction negatively regulates T-cell receptor signaling. Interacts with FCRL3. Interacts with VAV1. Interacts with CBL; this interaction promotes ubiquitination, internalization and subsequent degradation of CD247/CD3Z. Identified in a complex with CBL and UBE2L3. Interacts with SHB. Interacts with adapter protein SLA2; this interaction negatively regulates T-cell receptor signaling. Interacts with CBLB. Interacts (via SH2 domains) with RHOH; this interaction regulates ZAP70 subcellular localization. Interacts with DEF6 (By similarity). Interacts (ubiquitinated form) with OTUD7B and UBASH3B.

Family&Domains:

Composed of 2 N-terminal SH2 domains and a C-terminal kinase domain. The tandem SH2 domains bind to the doubly phosphorylated tyrosine-based activation motif (ITAM) of CD247/CD3Z and the non-canonical phosphorylated tyrosine-based activation motif (TAM) of RHOH (By similarity). The interdomain B located between the second SH2 and the kinase domain contains 3 tyrosines (Tyr-292, Tyr-315, Tyr-319) that are phosphorylated following TCR activation. These sites have been implicated in binding to other signaling molecules including CBL or VAV1. Thus, ZAP70 can also function as a scaffold by recruiting additional factors to the stimulated TCR complex.

Belongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily.

Research Fields

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

· Human Diseases > Immune diseases > Primary immunodeficiency.

· 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). Zhang J et al. Regulation of T Cell Activities in Rheumatoid Arthritis by the Novel Fusion Protein IgD-Fc-Ig. Frontiers in Immunology 2020 May 15;11:755. (PubMed: 32499775) [IF=7.3]

Application: WB    Species: mouse    Sample: spleen

FIGURE 9 | Effects of IgD-Fc-Ig (DG) on the protein expression of Lck, p-Lck, ZAP70 and p-ZAP70. (A–C) Western blot analysis of p-Lck and p-ZAP70 expression in PBMCs of RA patients, PBMCs were treated with IgD (3µg/mL) and IgD-Fc-Ig (1, 3, 10µg/mL) for 48h and lysed. (D–F) Western blot analysis of Lck, p-Lck, ZAP70 and p-ZAP70 expression in CIA mice. Mice spleens from each group were isolated and homogenized in lysis buffer.

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