ZAP70 antibody

Synonyms:Tyrosine-protein kinase ZAP-70|70 kDa zeta-chain associated protein|Syk-related tyrosine kinase|ZAP70|SRK antibody
Catalogue No.:FNab09583Reactivity:Human, Mouse
Host:RabbitTested Application:ELISA, IHC, WB, FC, IP, IF
Clonality:polyclonalIsotype:IgG
Size Price
100µg Inquiry
Dispatch Time: About 3 working days
  • SPECIFICATIONS
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Product Name
ZAP70 antibody
Catalogue No.
FNab09583
Size
100μg
Form
liquid
Purification
Immunogen affinity purified
Purity
≥95% as determined by SDS-PAGE
Clonality
polyclonal
Isotype
IgG
Storage
PBS with 0.02% sodium azide and 50% glycerol pH 7.3, -20℃ for 12 months(Avoid repeated freeze / thaw cycles.)
Immunogen
Immunogen
zeta-chain(TCR) associated protein kinase 70kDa
Alternative Names
Tyrosine-protein kinase ZAP-70|70 kDa zeta-chain associated protein|Syk-related tyrosine kinase|ZAP70|SRK antibody
UniProt ID
P43403
Observed MW
70 kDa
Application
Tested Applications
ELISA, IHC, WB, FC, IP, IF
Recommended dilution
WB: 1:1000-1:10000; IHC: 1:20-1:200; IP: 1:500-1:5000; IF: 1:20-1:200
Validated Images
Jurkat cells were subjected to SDS PAGE followed by western blot with FNab09583(ZAP70 Antibody) at dilution of 1:3000
IP Result of anti-ZAP70 (IP:FNab09583, 4ug; Detection:FNab09583 1:1000) with Jurkat cells lysate 3200ug.
Immunohistochemistry of paraffin-embedded human lymphoma using FNab09583(ZAP70 antibody) at dilution of 1:50
Background
Tyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates motility, adhesion and cytokine expression of mature T-cells, as well as thymocyte development. Contributes also to the development and activation of primary B-lymphocytes. When antigen presenting cells(APC) activate T-cell receptor(TCR), a serie of phosphorylations lead to the recruitment of ZAP70 to the doubly phosphorylated TCR component CD247/CD3Z through ITAM motif at the plasma membrane. This recruitment serves to localization to the stimulated TCR and to relieve its autoinhibited conformation. Release of ZAP70 active conformation is further stabilized by phosphorylation mediated by LCK. Subsequently, ZAP70 phosphorylates at least 2 essential adapter proteins: LAT and LCP2. In turn, a large number of signaling molecules are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Furthermore, ZAP70 controls cytoskeleton modifications, adhesion and mobility of T-lymphocytes, thus ensuring correct delivery of effectors to the APC. ZAP70 is also required for TCR-CD247/CD3Z internalization and degradation through interaction with the E3 ubiquitin-protein ligase CBL and adapter proteins SLA and SLA2. Thus, ZAP70 regulates both T-cell activation switch on and switch off by modulating TCR expression at the T-cell surface. During thymocyte development, ZAP70 promotes survival and cell-cycle progression of developing thymocytes before positive selection(when cells are still CD4/CD8 double negative). Additionally, ZAP70-dependent signaling pathway may also contribute to primary B-cells formation and activation through B-cell receptor(BCR).
How many times can antibodies be recycled?

First, usually it's not suggested to recycle antibodies. After use, buffer system of antibodies has changed. The storage condition of recycled antibodies for different customers also varies. Thus, the performance efficiency of recycled antibodies can’t be guaranteed. Besides, FineTest ever conducted the antibody recycling assay. Assay results show recycling times of different antibodies also varies. Usually, higher antibody titer allows more repeated use. Customers can determine based on experimental requirements.

Notes: After incubation, we recycle rest antibodies to centrifuge tube and store at 4℃. High titer antibodies can be stored for a minimum of one week. Reuse about three times.

What are components of FineTest antibody buffer?

Components of FineTest antibody buffer are usually PBS with proclin300 or sodium azide, BSA, 50% glycerol. Common preservative is proclin300 or sodium azide, which is widely applied in the lab and industry.

How about the storage temperature and duration of FineTest antibodies?

Most antibodies are stored at -20℃. Directly-labeled flow cytometry antibodies should be stored at 2 - 8℃. The shelf life is one year. If after sales issues for purchased antibodies appear, return or replacement is available. Usually, antibodies can be still used after the one-year warranty. We can offer technical support services.

Is dilution required for FineTest antibodies? What’s the dilute solution?

Directly-labeled flow cytometry antibodies are ready-to-use without dilution. Other antibodies are usually concentrated. Follow the dilution ratio suggested in the manual. Dilute solution for different experiments also varies. Common antibody dilution buffers are acceptable(e.g. PBST, TBST, antibody blocking buffer).

How to retrieve antibodies for immunohistochemistry?

Common retrieval buffers: Tris-EDTA Buffer(pH 9.0); Citrate Buffer(pH 6.0)

Heat induced antibody retrieval:

Method 1: Water-bath heating: Put the beaker with retrieval buffer and slide in the boiling water bath. Keep the boiling state for 15min. Naturally cool to room temperature;

Method 2: Microwave retrieval: Put the beaker with retrieval buffer and slide in the microwave oven. Heat at high power for 5min, Switch OFF for 3min, Heat at medium power for 5min. Naturally cool to room temperature.

How to choose secondary antibodies?

(1) Secondary antibodies react with primary antibodies. Thus, secondary antibodies should be against host species of primary antibodies. E.g. If the primary antibody is derived from rabbit, the relevant secondary antibody should be against rabbit. E.g. goat anti rabbit or donkey anti rabbit.

(2) Choose secondary antibody conjugates according to the experimental type, e.g. ELISA, WB, IHC etc. Common enzyme conjugated secondary antibodies are labelled by HRP, AP etc. Fluorescin or dye labelled secondary antibodies are applied in immunofluorescence and flow cytometry(e.g. FITC, Cy3).

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