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ZAP70 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-08469

Novus Biologicals, part of Bio-Techne
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NBP2-08469

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for ZAP70 Overexpression Lysate

ZAP70 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC208838

Accession#: NM_001079

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

69.7 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.

Type

Overexpression

Scientific Data Images for ZAP70 Overexpression Lysate

Western Blot: ZAP70 Overexpression Lysate [NBP2-08469]

Western Blot: ZAP70 Overexpression Lysate [NBP2-08469]

Western Blot: ZAP70 Overexpression Lysate (Adult Normal) [NBP2-08469] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ZAP70.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

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

Additional ZAP70 Products

Product Documents for ZAP70 Overexpression Lysate

Certificate of Analysis

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

Product Specific Notices for ZAP70 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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