Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Simple Western

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all ZAP70 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human ZAP70
Trp163-Cys254
Accession # P43403

Specificity

Detects human ZAP70.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human ZAP70 Antibody

Detection of Human ZAP70 antibody by Western Blot.

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.

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

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.

Detection of Human ZAP70 antibody by Simple WesternTM.

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.

Detection of Human ZAP70 by Simple Western

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.

Applications for Human ZAP70 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymphoma subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Simple Western

10 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and MOLT‑4 human acute lymphoblastic leukemia cell line

Western Blot

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line, and MOLT-4 human acute lymphoblastic leukemia cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

mrtle, mur, SRK, STD, TZK

Entrez Gene IDs

7535 (Human); 22637 (Mouse); 301348 (Rat)

Gene Symbol

ZAP70

UniProt

P43403

Additional ZAP70 Products

Product Documents for Human ZAP70 Antibody

Certificate of Analysis

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Product Specific Notices for Human ZAP70 Antibody

For research use only

Citations for Human ZAP70 Antibody

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Protocols

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