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ZAP70 Antibody (2F3.2)

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-44502

Novus Biologicals, part of Bio-Techne
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NBP2-44502-0.02mg
NBP2-44502-0.1mg

Key Product Details

Species Reactivity

Validated:

Human

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2a Kappa Clone # 2F3.2

Concentration

0.2 mg/ml

Product Summary for ZAP70 Antibody (2F3.2)

Immunogen

Recombinant ZAP-70 protein including residues 1-254 and encompassing SH2 domains of human ZAP70 (Uniprot: P43403)

Localization

Cell Surface and Cytoplasmic

Specificity

ZAP70 is a 70kDa protein tyrosine kinase found in T-cells and natural killer cells. Control of this protein translation is via the IgVH gene. In Western blotting of whole cell lysates of normal peripheral blood mononuclear cells, the antibody labels a band corresponding to ZAP70. In Western blotting of whole cell lysates of CD19-positive Purified leukemia cells from patients with Ig-unmutated and Ig-mutated CLL, the antibody labels a band corresponding to ZAP70 in the Ig-unmutated CLL samples, whereas no band is observed in the Ig-mutated CLL samples. In Western blotting of cell lysates of Jurkat cells (T-lymphoblastic cell line), the antibody labels a band of 70kDa protein. In Western blotting of cell lysates of A431 cells (carcinoma cell line), no band is observed. ZAP70 protein is expressed in leukemic cells of approximately 25% of chronic lymphocytic leukemia (CLL) cases as well. Anti-ZAP70 expression is an excellent surrogate marker for the distinction between the Ig-mutated (anti-ZAP70 negative) and Ig-unmutated (anti-ZAP70 positive) CLL subtypes and can identify patient groups with divergent clinical courses. The anti-ZAP70 positive Ig-unmutated CLL cases have been shown to have a poorer prognosis.

Marker

Chronic Lymphocytic Leukemia Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2a Kappa

Theoretical MW

70 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-47774)

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.

Scientific Data Images for ZAP70 Antibody (2F3.2)

Immunocytochemistry/ Immunofluorescence: ZAP70 Antibody (2F3.2) [NBP2-44502]

Immunocytochemistry/ Immunofluorescence: ZAP70 Antibody (2F3.2) [NBP2-44502]

Immunocytochemistry/Immunofluorescence: ZAP70 Antibody (2F3.2) [NBP2-44502] - Immunofluorescence Analysis of PFA-fixed Jurkat cells labeled with ZAP70 Antibody (2F3.2) followed by Goat anti-Mouse IgG-CF488 (Green). The nuclear counterstain is Nucspot (Red).
Immunohistochemistry-Paraffin: ZAP70 Antibody (2F3.2) [NBP2-44502]

Immunohistochemistry-Paraffin: ZAP70 Antibody (2F3.2) [NBP2-44502]

Immunohistochemistry-Paraffin: ZAP70 Antibody (2F3.2) [NBP2-44502] - Human Tonsil stained with ZAP70 Monoclonal Antibody (2F3.2).
Flow Cytometry: ZAP70 Antibody (2F3.2) [NBP2-44502]

Flow Cytometry: ZAP70 Antibody (2F3.2) [NBP2-44502]

Flow Cytometry: ZAP70 Antibody (2F3.2) [NBP2-44502] - Flow Cytometric Analysis of PFA-fixed Jurkat cells. ZAP70 Antibody (2F3.2) followed by goat anti-Mouse IgG-CF488 (Blue); Isotype Control (Red).

Applications for ZAP70 Antibody (2F3.2)

Application
Recommended Usage

Flow Cytometry

1-2 ug/million cells

Immunocytochemistry/ Immunofluorescence

1-2 ug/ml

Immunohistochemistry-Paraffin

1-2 ug/ml

Simple Western

10 ug/ml
Application Notes
Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
Optimal dilution for a specific application should be determined.
In Simple Western only 10 - 15 ul of the recommended dilution is used per data point. Separated by Size.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10 mM PBS with 0.05% BSA

Preservative

0.05% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C.

Background: ZAP70

Zap-70, a Syk-family protein tyrosine kinase, plays a critical role in mediating T cell signal transduction in response to T cell receptor (TCR) activation (1). TCR-mediated activation of the Src-family kinases, Lck and Fyn, results in tyrosine phosphorylation of the TCR zeta and CD3 chains. These domains serve as targets for binding of ZAP-70 via its tandem SH2 domains. This binding correlates with activation of ZAP-70, a critical event in T cell activation (2). Following TCR engagement, ZAP-70 is phosphorylated on several tyrosine residues, presumably by two mechanisms: an autophosphorylation and a trans-phosphorylation by the Src-family tyrosine kinase, Lck. Phosphorylation of Tyr319 is required for full activation and increased positive downstream regulation by ZAP-70 (3).

Long Name

Zeta-chain [TCR] Associated Protein Kinase 70 kDa

Alternate Names

mrtle, mur, SRK, STD, TZK

Gene Symbol

ZAP70

UniProt

Additional ZAP70 Products

Product Documents for ZAP70 Antibody (2F3.2)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for ZAP70 Antibody (2F3.2)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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