Human ZAP70 Antibody

Catalog # Availability Size / Price Qty
AF3709
AF3709-SP
Detection of Human ZAP70 by Western Blot.
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Product Details
Citations (1)
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Human ZAP70 Antibody Summary

Species Reactivity
Human
Specificity
Detects human ZAP70.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human ZAP70
Trp163-Cys254
Accession # P43403
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human ZAP70 antibody by Western Blot. View Larger

Detection of Human ZAP70 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MOLT‑4 human acute lymphoblastic leukemia cell line, Daudi human Burkitt's lymphoma cell line (negative control), and Raji human Burkitt's lymphoma cell line (negative control). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human ZAP70 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3709) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for ZAP70 at approximately 70 kDa (as indicated). GAPDH is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Immunohistochemistry ZAP70 antibody in Human Lymphoma by Immunohistochemistry (IHC-P). View Larger

ZAP70 in Human Lymphoma. ZAP70 was detected in immersion fixed paraffin-embedded sections of human lymphoma using 10 µg/mL Human ZAP70 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3709) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western Detection of Human ZAP70 antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human ZAP70 by Simple WesternTM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, MOLT-4 human acute lymphoblastic leukemia cell line, Ramos human Burkitt's lymphoma cell line, and Raji human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for ZAP70 at approximately 66 and 149 kDa (as indicated) using 10 µg/mL of Goat Anti-Human ZAP70 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3709) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Simple Western Detection of Human ZAP70 by Simple Western View Larger

Detection of Human ZAP70 by Simple Western TIM-3 Association with Intracellular Kinases in CD8+/MART-1+ T cells.TIM-3 co-immunoprecipitation analysis of unactivated and 15 min. stimulation with anti-CD3/CD28 beads (activated). Equivalent amounts of protein (~2mg) were co-immunoprecipitated with pAb anti-TIM-3 antibody and western blot was performed using capillary electrophoresis. Cleared lysate served as a loading control for individual antibody reactivity. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0140694), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ZAP70

ZAP70 (zeta-chain (TCR) associated protein kinase 70 kDa), expressed primarily in T and NK cells, is a Syk family cytosolic protein tyrosine kinase that consists of two N-terminal SH2 domains and a C-terminal tyrosine kinase domain. Upon T cell receptor activation and phosphorylation of TCR ITAMs by Src family kinases, ZAP70 is recruited to phosphorylated ITAM sequences and subsequently phosphorylated on several tyrosine residues. ZAP70 has been implicated in several immune disorders. An autosomal recessive form of SCID in humans has been attributed to a homozygous mutation in the kinase domain of ZAP70. ZAP70 expression also defines an aggressive subset of CLL.

Long Name
Zeta-chain [TCR] Associated Protein Kinase 70 kDa
Entrez Gene IDs
7535 (Human); 22637 (Mouse); 301348 (Rat)
Alternate Names
70 kDa zeta-associated protein; EC 2.7.10; EC 2.7.10.2; mrtle; mur; SRK; SRKFLJ17679; STD; STDFLJ17670; Syk-related tyrosine kinase; tyrosine-protein kinase ZAP-70; TZK; ZAP70; ZAP-70; zeta-chain (TCR) associated protein kinase (70 kD); zeta-chain (TCR) associated protein kinase 70kDa; zeta-chain associated protein kinase, 70kD

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Citation for Human ZAP70 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. TIM-3 Suppresses Anti-CD3/CD28-Induced TCR Activation and IL-2 Expression through the NFAT Signaling Pathway.
    Authors: Tomkowicz B, Walsh E et al.
    PLoS One
    Species: Human
    Sample Types: blood
    Applications: Simple Western

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