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NOXO1 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-13735

Novus Biologicals, part of Bio-Techne
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NBL1-13735

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for NOXO1 Overexpression Lysate

NOXO1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC204220

Accession#: NM_144603

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

40.5 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 NOXO1 Overexpression Lysate

Western Blot: NOXO1 Overexpression Lysate [NBL1-13735]

Western Blot: NOXO1 Overexpression Lysate [NBL1-13735]

Western Blot: NOXO1 Overexpression Lysate (Adult Normal) [NBL1-13735] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NOXO1.

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: NOXO1

NOXO1 (Nox organizing protein 1) and NOXA1 (Nox Activating protein 1) are homologs of p47phox and p67phox. p47phox functions in phagocytes as an essential organizing protein mediating the binding of other regulatory proteins during activation of the phagocyte oxidase, and its translocation to the membrane is triggered upon cell activation by hyperphosphorylation, which relieves autoinhibition of SH3 and PX domains. NOXO1 lacks an auto inhibitory region and phosphorylation sites that are present in p47phox. Co-transfection of Nox1, NOXO1 and NOXA1 reconstitutes ROS (reactive oxygen species) generation in HEK 293 cells in the absence of cell stimulation. NOXO1 binds to the phosphatidylinositol (PtdIns) lipids PtdIns 3,5-P2, PtdIns 5-P and PtdIns 4-P. Unlike p47phox, which is located in the cytosol of resting cells and translocates to the plasma membrane where gp91phox is located, NOXO1 co-localizes with Nox1 in the membranes of resting cells. This localization of NOXO1 is dictated by its PX domain, since this domain but not the remainder of the molecule localizes to membranes.

Alternate Names

MGC20258, NADPH oxidase organizer 1, Nox organizer 1, Nox-organizing protein 1, P41NOX, P41NOXA, P41NOXB, P41NOXC, regulatory protein P41NOX, SH3 and PX domain-containing protein 5, SH3PXD5NADPH oxidase regulatory protein, SNX28

Gene Symbol

NOXO1

Additional NOXO1 Products

Product Documents for NOXO1 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 NOXO1 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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