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AM03112PU-N  ZAP70 antibody

see related secondary antibodies
see all 43 ZAP70 products
0.1 mg / US$ 295
NOVUS BIOLOGICALS
PO Box 802 Littleton, CO 80160, USA
E-Mail: novus@novus-biologicals.com
Homepage: http://www.novus-biologicals.com

Quick Overview

Mouse anti ZAP70 ZAP-03

Synonyms

ZAP-70; ZAP 70; Tyrosine-protein kinase ZAP-70; Syk-related tyrosine kinase; SRK

Product Description

Mouse anti ZAP70 ZAP-03, Presentation: Aff - Purified. Product is tested for Flow cytometry ( F ), Immunofluorescence ( IF ), Westernblot / Immunoblot ( WB )

Properties

ApplicationsFlow cytometry ( F ), Immunofluorescence ( IF ), Westernblot / Immunoblot ( WB )
ReactivityHuman ( Hu )
PresentationAff - Purified
HostMouse
IsotypeIgG1
CloneZAP-03
Catalog NumberAM03112PU-N
Swiss Prot. No.P43403
Quantity0.1 mg
PriceUS$ 295
DeliveryWorldwide
ManufacturerAcris Antibodies GmbH
Datasheetview PDF-Download
AM03112PU-N.pdf

Datasheet Extract

Monoclonal Antibody to ZAP70
Concentration1.0 mg/ml
BackgroundThe ZAP70 (zeta-associated protein of 70 kDa) tyrosine kinase was identified as a tyrosine phosphoprotein that associates with TCR zeta subunit and undergoes tyrosine phosphorylation following TCR stimulation. ZAP70 is a Syk family tyrosine kinase primarily expressed in T and NK cells that plays an essential role in signaling through the TCR. TCR-mediated activation of T cells is crucial to the immune response. In humans, ZAP70 gene mutations resulting in lower ZAP70 protein expression levels or expression of catalytically inactive ZAP70 proteins, have been identified. ZAP70 deficiency results in the absence of mature CD8+ T cells and the prevention of TCR-mediated activation of CD4+ T cells, and it can lead to severe combined immunodeficiency. In patients with chronic lymphocytic leukemia (B-CLL), ZAP70 expression on B cell was shown to be correlated with disease progression and survival.
ZAP70 is cytosolic protein migrating at 70 kDa in SDS-PAGE. It contains two N-terminal SH2 domains (Src homology domain 2) and a C-terminal kinase domain. Crystal structure of the ZAP70 SH2 domains in complex with a TCR zeta subunit peptide was described. During T cell activation , the binding of ZAP70 SH2 domains to the phosphorylated zeta subunit on the activated TCR complex causes a colocalization with the Lck tyrosine kinase that phosphorylates ZAP70 on Tyr493 in the activation loop. ZAP70 autophosphorylates multiple tyrosines in the region between the SH2 domains and the kinase domain, including the binding sites for additional SH2-containing signaling proteins such as SLP76, LAT, Lck, PLCgamma1, Vav, Shc, Ras-GAP, and Abl. ZAP70-mediated activation of these downstream effectors leads to the release of intracellular calcium stores, and the transcription of interleukin-2 and other genes important for an immune response.
ImmunogenBacterially expressed fusion protein representing C-terminal part (160 amino acids) of human ZAP-70.
Swiss Num.: P43403
Format
State: Liquid purified IgG fraction (> 95% pure by SDS-PAGE).
Purification: Protein A affinity chromatography.
BufferSystem: PBS, pH 7.4 containing 15 mM sodium azide as preservative.
ApplicationsFlow Cytometry (2-5 μg/ml):Intracellular staining.
Positive control: HPB-ALL human peripheral blood T cell leukemia cell line.
Western Blotting (0,5 μg/ml):Under Reducing conditions. SDS-PAGE (10% separating gel).
Positive control: HPB-ALL human peripheral blood T cell leukemia cell line.
Negative control: RAMOS human Burkitt lymphoma cell line.
Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate (1:1) with reducing Laemmli SDS-PAGE sample buffer. Boil for 5 min.
Immunocytochemistry.
SpecificityThe antibody ZAP-03 reacts with ZAP70, a 70 kDa protein tyrosine kinase expressed in T and NK cells.
ZAP70 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.
Species: Human.
StorageStore the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
References1. Ishaq M, DeGray G, Natarajan V: Evidence for the involvement of tyrosine kinase ZAP 70 in nuclear retinoid receptor-dependent transactivation in T lymphocytes. J Biol Chem. 2005 Oct 7;280(40):34152-8.
2. Schneider H, Smith X, Liu H, Bismuth G, Rudd CE: CTLA-4 disrupts ZAP70 microcluster formation with reduced T cell/APC dwell times and calcium mobilization. Eur J Immunol. 2008 Jan;38(1):40-7.
3. Chan AC, Irving BA, Fraser JD, Weiss A.: The zeta chain is associated with a tyrosine kinase and upon T-cell antigen receptor stimulation associates with ZAP-70, a 70-kDa tyrosine phosphoprotein. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9166-70.

14 Secondary Antibodies

Catalog No.Name / HostPresentationReactivity 
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R1253F Mouse IgG (H&L)  
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R1253T Mouse IgG (H&L)  
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R1253TR Mouse IgG (H&L)  
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R1253HRP Mouse IgG (H&L)  
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R1253AP Mouse IgG (H&L)  
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  Goat Cy2   1 mg / US$ 445
   
R1457C3 Mouse IgG (H&L) multi-species ads.  
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R1457C35 Mouse IgG (H&L) multi-species ads.  
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R1457C5 Mouse IgG (H&L) multi-species ads.  
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R1457C55 Mouse IgG (H&L) multi-species ads.  
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R1405R Mouse IgG (H&L) F(ab')2 Fragment ads. to Bov, Eq, Hu, Rb, Rt, Sh  
  Goat PE   1 ml / US$ 455
   
R1403R Mouse IgG (H&L) F(ab')2 Fragment ads. to Hu serum proteins  
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R1455R Mouse IgG (H&L) F(ab')2 Fragment multi-species ads.  
  Donkey PE   1 ml / US$ 455
   

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