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ZAP70 Antibody (YP4051) (YA003)

(Synonyms: SRK, ZAP70, Tyrosine-protein kinase ZAP-70, 70 kDa zeta-chain associated protein, Syk-related tyrosine kinase)
Customer Review

Based on 1 Customer Validation

ZAP70 Antibody (YP4051) (YA003) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ZAP70.

For research use only. We do not sell to patients.
  • Host:

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, ICC/IF, IHC-P, FC, IP

  • Reactivity :

    Human

  • Formulation:

    Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

  • Conjugation:
    Non-conjugated
ALL WB FC
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • ZAP70 Antibody (YP4051) (YA003) Product Image
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To place orders, for customer services and technical support, please contact: MedChemExpress USA

Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Related Products:
Top Publications Citing

Use of MCE Products

Applications

Application
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
FC Info
FC: Flow Cytometry
IP Info
IP: Immunoprecipitation
Dilution Ratio 1:500-1:1000 1:50-1:200 1:50-1:200 1:50-1:100 1:10-1:50

Product Details

Description

ZAP70 Antibody (YP4051) (YA003) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ZAP70.

  • Host Rabbit
  • Clonality Recombinant,Monoclonal
  • Species Reactivity
    Human
  • Observed Molecular Weight
    Observed band size: 74 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 70 kDa
Species Reactivity Database

Entrez Gene: 7535 Human

SwissProt: P43403 Human

Immunogen

Synthetic peptide corresponding to Human ZAP70.AA range:570-619.

Sensitivity

Endogenous

Purification

Protein A affinity purified.

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG

RRID

AB_3102594

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
    Western blot analysis of extracts from Jurkat(lane 2(20μg) ) and Jurkat(lane 3(40μg) ) using ZAP70 (HY-P80524) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80524, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
    Western blot analysis was performed on protein extracts from Jurkat (lane 2, 20 μg), and Jurkat (lane 3, 40 μg) using ZAP70 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
    Flow cytometric analysis of 1X106 Jurkat cells labeling ZAP70 Antibody (HY-P80524, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Background

  • Function

    ZAP70 (zeta-chain-associated protein kinase 70) is a cytoplasmic protein tyrosine kinase that functions as a central signaling mediator downstream of the T-cell receptor (TCR) and is required for the initiation of antigen receptor-driven T-cell responses[1]. Mechanistically, TCR engagement leads to phosphorylation of immunoreceptor tyrosine-based activation motifs (ITAMs), recruitment of ZAP70 through its tandem SH2 domains, and subsequent phosphorylation of adaptor proteins including LAT and SLP-76, thereby propagating signaling pathways that regulate T-cell activation, differentiation, proliferation, and cytokine production[1][5]. Because of this pivotal role in adaptive immunity, ZAP70 is essential for normal thymocyte development as well as peripheral T-cell function[1][5]. In disease settings, loss-of-function mutations in ZAP70 cause a combined immunodeficiency characterized by profound defects in T-cell signaling, markedly reduced CD8+ T-cell populations, recurrent infections, and immune dysregulation[2][3]. Altered ZAP70 signaling has also been linked to autoimmune phenotypes, highlighting the requirement for tightly regulated TCR signaling thresholds during immune homeostasis[2]. Compared with the closely related kinase SYK, ZAP70 exhibits a specialized role in T-cell receptor signaling, whereas SYK displays broader expression and higher intrinsic kinase activity in multiple immune cell types[6]. For experimental research, ZAP70 serves as a widely used molecular node for dissecting TCR signal transduction networks and immune-cell activation mechanisms, and its kinase activity can be selectively inhibited by compounds such as epigallocatechin gallate (EGCG) in cellular studies[4].

  • Subcellular Localization

    Cytoplasm; Cell membrane; Peripheral membrane protein

  • Expression


    Tissue_specificity:It is expressed in T cells and natural killer cells. It is also present in early thymocytes and pre-B cells/pre-B cells.

  • Isoforms & Post-Translational Modification

    P43403 has 3 isomers: P43403-1: 69872 Da (predicted); P43403-2: 35647 Da (predicted); P43403-3: 55873 Da (predicted).
    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

  • Subunit

    Interacts with CD247/CD3Z; this interaction docks ZAP70 at the stimulated TCR (PubMed:1423621, PubMed:26783323, PubMed:7659156). Interacts with NFAM1 (PubMed:15143214).

  • SwissProt ID

    P43403

  • Gene ID
    7535 [NCBI]
  • Synonyms

    SRK, ZAP70, Tyrosine-protein kinase ZAP-70, 70 kDa zeta-chain associated protein, Syk-related tyrosine kinase

  • Research Field

    Immunology

References

[1]. Noraz N, et al. Alternative antigen receptor (TCR) signaling in T cells derived from ZAP-70-deficient patients expressing high levels of Syk. J Biol Chem. 2000;275(21):15832-15838.

[2]. Fernández-Aguilar LM, et al. A Story of Kinases and Adaptors: The Role of Lck, ZAP-70 and LAT in Switch Panel Governing T-Cell Development and Activation. Biology (Basel). 2023;12(9):1163.

[3]. Ashouri JF, et al. ZAP70, too little, too much can lead to autoimmunity. Immunol Rev. 2022 May;307(1):145-160. [Content Brief]

[4]. Sharifinejad N, et al. Clinical, Immunological, and Genetic Features in 49 Patients With ZAP-70 Deficiency: A Systematic Review. Front Immunol. 2020 May 5;11:831. [Content Brief]

[5]. Wang H, et al. ZAP-70: an essential kinase in T-cell signaling. Cold Spring Harb Perspect Biol. 2010;2(5):a002279. [Content Brief]

[6]. ZAP70 gene information from NCBI.

[7]. Wang H, et al. ZAP-70: an essential kinase in T-cell signaling. Cold Spring Harb Perspect Biol. 2010 May;2(5):a002279. [Content Brief]

[8]. Zhang W, et al. LAT: the ZAP-70 tyrosine kinase substrate that links T cell receptor to cellular activation. Cell. 1998 Jan 9;92(1):83-92. [Content Brief]

[9]. Arpaia E, et al. Defective T cell receptor signaling and CD8+ thymic selection in humans lacking zap-70 kinase. Cell. 1994 Mar 11;76(5):947-58. [Content Brief]

[10]. Wiestner A, et al. ZAP-70 expression identifies a chronic lymphocytic leukemia subtype with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. Blood. 2003 Jun 15;101(12):4944-51. [Content Brief]

[11]. Noraz N, et al. Alternative antigen receptor (TCR) signaling in T cells derived from ZAP-70-deficient patients expressing high levels of Syk. J Biol Chem. 2000 May 26;275(21):15832-8. [Content Brief]

[12]. Visperas PR, et al. Identification of Inhibitors of the Association of ZAP-70 with the T Cell Receptor by High-Throughput Screen. SLAS Discov. 2017 Mar;22(3):324-331. [Content Brief]

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ZAP70 Antibody (YP4051) (YA003) Related Classifications

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     Western blot analysis of extracts from Jurkat(lane 2(20μg) ) and Jurkat(lane 3(40μg) ) using ZAP70 (HY-P80524) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80524, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

  • 1/16

    MedChemExpress Validation
     Western blot analysis was performed on protein extracts from Jurkat (lane 2, 20 μg), and Jurkat (lane 3, 40 μg) using ZAP70 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • 1/16

    MedChemExpress Validation
     Flow cytometric analysis of 1X106 Jurkat cells labeling ZAP70 Antibody (HY-P80524, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • ZAP70 Antibody (YP4051) (YA003) Product Image
    1/16
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • ZAP70 Antibody (YP4051) (YA003) Product Image
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
    1/1
    MedChemExpress Validation
     Western blot analysis of extracts from Jurkat(lane 2(20μg) ) and Jurkat(lane 3(40μg) ) using ZAP70 (HY-P80524) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80524, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
    1/1
    MedChemExpress Validation
     Western blot analysis was performed on protein extracts from Jurkat (lane 2, 20 μg), and Jurkat (lane 3, 40 μg) using ZAP70 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
    1/1
    MedChemExpress Validation
     Flow cytometric analysis of 1X106 Jurkat cells labeling ZAP70 Antibody (HY-P80524, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • Experimental Validation Results for ZAP70 Antibody (YP4051) (YA003)
  • ZAP70 Antibody (YP4051) (YA003) Product Image
    MedChemExpress Validation

    Western blot analysis of extracts from Jurkat(lane 2(20μg) ) and Jurkat(lane 3(40μg) ) using ZAP70 (HY-P80524) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80524, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

  • ZAP70 Antibody (YP4051) (YA003) Product Image
    MedChemExpress Validation

    Western blot analysis was performed on protein extracts from Jurkat (lane 2, 20 μg), and Jurkat (lane 3, 40 μg) using ZAP70 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • ZAP70 Antibody (YP4051) (YA003) Product Image
    MedChemExpress Validation

    Flow cytometric analysis of 1X106 Jurkat cells labeling ZAP70 Antibody (HY-P80524, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • ZAP70 Antibody (YP4051) (YA003) Product Image WB
  • ZAP70 Antibody (YP4051) (YA003) Product Image WB
  • ZAP70 Antibody (YP4051) (YA003) Product Image FC