GSK-3 beta Overexpression Lysate

Novus Biologicals | Catalog # NBL1-11362

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for GSK-3 beta Overexpression Lysate

GSK-3 beta Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC200468

Accession#: NM_002093

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.
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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.

Theoretical MW

47.9 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 GSK-3 beta Overexpression Lysate

Western Blot: GSK-3 beta Overexpression Lysate [NBL1-11362]

Western Blot: GSK-3 beta Overexpression Lysate [NBL1-11362]

Western Blot: GSK3 beta Overexpression Lysate (Adult Normal) [NBL1-11362] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GSK3 beta.

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: GSK-3 beta

Glycogen synthase kinase 3beta (GSK-3 beta) is a Serine/Threonine protein kinas involved in the insulin and wingless pathways (1-2). Originally identified as a regulator of glycogen synthase, it has been shown to regulate a diverse array of cellular functions. GSK-3 beta activity is down regulated by phosphorylation on Serine 9 by PKB, or activated by phosphorylation on Tyrosine 216. Once activated, the protein can phosphorylate p53, c-jun, heat shock factor-1 and cyclin D1 (3-4). It is also known to phosphorylate Tau in Allzheimer's disease. Additionally, increased GSK-3 beta protein levels are found in Alzheimer's disease brains.

Long Name

Glycogen Synthase Kinase 3 beta

Alternate Names

GSK3 beta, GSK3B

Gene Symbol

GSK3B

Additional GSK-3 beta Products

Product Documents for GSK-3 beta Overexpression Lysate

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 GSK-3 beta 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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