Key Product Details
Species Reactivity
Human
Applications
Immunohistochemistry, Immunohistochemistry-Paraffin, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2a Kappa Clone # 2F3.2
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Product Specifications
Immunogen
Recombinant ZAP-70 protein including residues 1-254 and encompassing SH2 domains of human ZAP70 (Uniprot: P43403)
Localization
Cell Surface and Cytoplasmic
Marker
Chronic Lymphocytic Leukemia Marker
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.
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.
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.
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] - 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] - 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 Cytometric Analysis of PFA-fixed Jurkat cells. ZAP70 Antibody (2F3.2) followed by goat anti-Mouse IgG-CF488 (Blue); Isotype Control (Red).Simple Western: ZAP70 Antibody (2F3.2) [NBP2-44502]
Simple Western: ZAP70 Antibody (2F3.2) [NBP2-44502] - Simple Western lane view shows a specific band for ZAP70 in 0.2 mg/ml of Jurkat (left) and H. Tonsil (right) lysate(s). This experiment was performed under reducing conditions using the 12-230 kDa separation system.Simple Western: ZAP70 Antibody (2F3.2) [NBP2-44502]
Simple Western: ZAP70 Antibody (2F3.2) [NBP2-44502] - Electropherogram image of the corresponding Simple Western lane. ZAP70 antibody was used at 10 ug/ml dilution of Jurkat and H. Tonsil lysates(s) respectively.Simple Western: ZAP70 Antibody (2F3.2) [NBP2-44502] -
Simple Western: ZAP70 Antibody (2F3.2) [NBP2-44502] - Lane view shows a specific band for ZAP70 in Jurkat cell lysate using 50 ug/mL antibody dilution. Electropherogram image of corresponding Simple Western lane view at WES molecular weight of 70 kDa. Image from an internal validation.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.
See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat and H. Tonsil lysates, separated by Size, antibody dilution of 10 ug/mL, apparent MW was 67 kDa.
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.
See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat and H. Tonsil lysates, separated by Size, antibody dilution of 10 ug/mL, apparent MW was 67 kDa.
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
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
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 or batch 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.
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Liperfluo
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Immunohistochemistry
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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