Overview

  • Product name
    Human ZAP70 ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Jurkat 5 4.7%
    Inter-assay
    Sample n Mean SD CV%
    Jurkat 3 6.5%
  • Sample type
    Cell culture extracts, Adherent cells, Suspension cells, Cell Lysate, Tissue Homogenate
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 500 pg/ml
  • Range
    1.6 ng/ml - 100 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture media 90 89% - 91%
    Fetal Bovine Serum 57 48% - 61%
    Bovine Serum Albumin 106 103% - 111%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Predicted to work with: RatDoes not react with: Mouse
  • Product overview

    Abcam’s ZAP70 Human in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of catalase protein in Human cell and tissue extracts.

    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    ZAP70 is a tyrosine kinase that plays an essential role in the regulation of the adaptive immune response. ZAP70 regulates motility, adhesion and cytokine expression of mature T-cells, as well as in thymocyte development. ZAP70 regulates T-cell activation by modulating T-Cell Receptor expression at the cell surface.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    10X ZAP70 Capture Antibody 1 x 600µl
    10X ZAP70 Detector Antibody 1 x 600µl
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
    ZAP70 Human Lyophilized Recombinant Protein 2 x 50ng
  • Research areas
  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily.
    Contains 1 protein kinase domain.
    Contains 2 SH2 domains.
  • Domain
    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.
  • Post-translational
    modifications
    Phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Tyr-319 phosphorylation is essential for full activity.
  • Cellular localization
    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.
  • Information by UniProt
  • Alternative names
    • 70 kDa zeta associated protein
    • 70 kDa zeta-associated protein
    • EC 2.7.10.2
    • FLJ17670
    • FLJ17679
    • Selective T cell defect
    • SRK
    • STD
    • Syk related tyrosine kinase
    • Syk-related tyrosine kinase
    • Truncated ZAP kinase
    • Tyrosine protein kinase ZAP70
    • Tyrosine-protein kinase ZAP-70
    • TZK
    • ZAP 70
    • ZAP70
    • ZAP70_HUMAN
    • Zeta chain associated protein kinase 70kD
    • Zeta chain associated protein kinase 70kDa
    • Zeta chain associated protein kinase 70kDa isoform 1
    • Zeta chain associated protein kinase 70kDa isoform 2
    • Zeta chain of T cell receptor associated protein kinase 70
    • Zeta chain TCR associated protein kinase 70kD
    • Zeta chain TCR associated protein kinase 70kDa
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab173192 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Example ZAP70 standard curve.  Background-subtracted data values (mean +/- SD) are graphed. Data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.

  • Titration of Jurkat extract within the working range of the Zap70 assay. Background subtracted data from duplicate measurements are plotted.

  • Interpolated values of ZAP70 are graphed for the indicated cells lines based on a extract load of 12.5 µg/mL. The ZAP70 detector antibody was used to analyze the same extracts by western blot (20 µg loaded/lane). The actin blot is included to show the relative loads of each lysate.

Protocols

References

ab173192 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab173192.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up