The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2 - 5 µg/ml. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 0.5 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
Plays a role in T-cell development and lymphocyte activation. Essential for TCR-mediated IL-2 production. Isoform 1 induces TCR-mediated signal transduction, isoform 2 does not.
Expressed in T- and natural killer cells.
Involvement in disease
Defects in ZAP70 are the cause of selective T-cell defect (STD) [MIM:176947]. STD is an autosomal recessive form of severe combined immunodeficiency characterized by a selective absence of CD8-type T-cells.
Belongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily. Contains 1 protein kinase domain. Contains 2 SH2 domains.
The SH2 domains bind to the phosphorylated tyrosine-based activation motif (TAM) of CD3Z and the non-canonical phosphorylated tyrosine-based activation motif (TAM) of RHOH.
Phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Tyr-319 phosphorylation is essential for full activity.
Cytoplasm. Cell membrane. After antigen stimulation, isoform 1 concentrates at the immunological synapse and isoform 2 remains cytoplasmic. Co-localizes together with RHOH in the immunological synapse. RHOH is required for its proper localization to the cell membrane and cytoskeleton fractions in the thymocytes.
Flow Cytometry analysis of the human peripheral blood of a patient with B-cell chronic lymphatic leukemia (B-CLL) using antibody ab38871; Dot Plot diagram shows the distribution of lymphocyte populations: T-lymphocytes (green), B-lymphocytes (blue) and NK cells (red).
Western blot - ZAP70 antibody [ZAP-03] (ab38871)
Lane 1 : Anti-ZAP70 antibody [ZAP-03] (ab38871) at 4 µg/ml Lane 2 : Anti-ZAP70 antibody [ZAP-03] (ab38871) at 2 µg/ml Lane 3 : Anti-ZAP70 antibody [ZAP-03] (ab38871) at 1 µg/ml Lane 4 : Anti-ZAP70 antibody [ZAP-03] (ab38871) at 0.5 µg/ml Lane 5 : ZAP-70 specific antibody 1E7.2 at 1 µg/ml Lane 6 : Isotypic control PPV-06 (mouse IgG1).
Lane 1 : HPB-ALL peripheral blood leukemia T-cell line Lane 2 : HPB-ALL peripheral blood leukemia T-cell line Lane 3 : HPB-ALL peripheral blood leukemia T-cell line Lane 4 : HPB-ALL peripheral blood leukemia T-cell line Lane 5 : HPB-ALL peripheral blood leukemia T-cell line Lane 6 : HPB-ALL peripheral blood leukemia T-cell line
Performed under reducing conditions.
Predicted band size : 70 kDa Observed band size : 70 kDa Additional bands at : 55 kDa (possible IgG,degradation product).
Western Blotting analysis of HPB-ALL peripheral blood leukemia T-cell line; SDS-PAGE; 10% separating gel, 4% stacking gel.
ZAP-70 is a molecule susceptible to degradation. It is recommended to use freshly prepared cell lysates (protease inhibitors are essential) to avoid non-specific staining of degradation products.