Daxx Overexpression Lysate

Name: Daxx Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-09622

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-09622

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Daxx Overexpression Lysate

Daxx Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC226603

Accession#: NM_001141969

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

81.2 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 Daxx Overexpression Lysate

Western Blot: Daxx Overexpression Lysate [NBP2-09622]

Western Blot: Daxx Overexpression Lysate (Adult Normal) [NBP2-09622] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Daxx.

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: Daxx

Daxx (death-domain-associated protein) was originally identified as a cytoplasmic protein that binds to the death domain of the transmembrane death receptor Fas. It is now that known that the a large porportion of Daxx molecules are nuclear and associate with the promyelocytic leukaemia nuclear body (PML-NB) and other subnuclear domains (reviewed in Salomoni and Khelifi). The promyelocytic leukemia protein Pml is an essential component of the PML-NB. Data suggests that certain stimuli including Fas stimulation, causes Daxx to be translocated from the nucleus to the cytoplasm. Daxx is ubiquitously expressed, and particularly high levels of Daxx have been reported in the thymus and testes. At the cellular level Daxx may be found in cytoplasm and within heterochromatic regions of the nucleus. Daxx has been reported to interact and co-localize with Pml in the nucleus of tumor cell lines and primary cells. Pml is necessary for Fas-induced cell death and Daxx pro-apoptotic functions. Daxx is thought to have both pro- and anti-apoptotic functions depending on the stimulus and the cell type. Daxx pro-apoptotic functions appear to occur in both the cytoplasm and nucleus. Although the mechanisms remain to be fully elucidated, research indicates that Daxx plays a role in the pathology of human diseases including cancer and neurodegerative disorders. Human Daxx is a 740 amino acid protein (GenBank no. gi/62898369).

Long Name

Fas Death Domain-associated Protein

Alternate Names

BING2, DAP6

Gene Symbol

DAXX

Additional Daxx Products

Product Documents for Daxx Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Daxx 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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