Glucokinase/GCK Overexpression Lysate

Name: Glucokinase/GCK Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-09947

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-09947

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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 Glucokinase/GCK Overexpression Lysate

Glucokinase/GCK Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC222793

Accession#: NM_033507

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

52 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 Glucokinase/GCK Overexpression Lysate

Western Blot: Glucokinase/GCK Overexpression Lysate [NBP2-09947]

Western Blot: GCK Overexpression Lysate (Adult Normal) [NBP2-09947] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GCK.

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: Glucokinase/GCK

Hexokinases phosphorylate hexose to form hexose 6-phosphate, the first step for hexose metabolism. There are four mammalian hexokinases (I, II, III and IV) and hexokinase IV is commonly known as glucokinase (GCK). Unlike hexokinase I, II and III, which have high affinity for glucose and are strongly inhibited by the product, glucose-6-phosphate, GCK has much lower affinity for glucose and is not inhibited by the product. Consequently, GCK has a Km for glucose of approximately 7 mM, which is 100 times higher than that of hexokinase I, II, and III. This unique enzymatic property of GCK allows it to respond to blood glucose levels and contribute to the maintenance of blood glucose levels within the normal physiological range of 4 mM to 6 mM. In the pancreatic islets, GCK serves as a glucose sensor to control insulin release in the beta cells, and to control glucagon release in the alpha cells. In hepatocytes, GCK responds to changes of ambient glucose levels by increasing or reducing glycogen synthesis. Mutations in GCK have been associated with non-insulin-dependent diabetes mellitus, maturity-onset diabetes of the young type 2, and hyperinsulinemia of infancy. The enzyme activity was measured using a phosphatase coupled kinase assay.

Long Name

Glucose Kinase

Alternate Names

FGQTL3, GCK, GLK, Hexokinase-4, Hexokinase-D, HHF3, HK4, HXKP, LGLK

Gene Symbol

GCK

Additional Glucokinase/GCK Products

Product Documents for Glucokinase/GCK Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Glucokinase/GCK 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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