ZAP70 Antibody [Unconjugated]

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.

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 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.

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 (//dx.plos.org/10.1371/journal.pone.0140694), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

ZAP70 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human ZAP70
  Trp163-Cys254
  Accession # P43403
• Specificity: Detects human ZAP70.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ZAP70
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ZAP70 Antibody [Unconjugated]

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ZAP70 Antibody (AF3709)(1)

Publication using AF3709

• Tomkowicz B, Walsh E et al. TIM-3 Suppresses Anti-CD3/CD28-Induced TCR Activation and IL-2 Expression through the NFAT Signaling Pathway. PLoS One 2024-10-22 [PMID: 26492563] (Simple Western, Human)

Bioinformatics

• Gene Symbol: ZAP70
• Uniprot:
  • P43403()