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N-Acetylmannosamine Kinase/GNE Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-07744

Novus Biologicals, part of Bio-Techne
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NBP2-07744

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for N-Acetylmannosamine Kinase/GNE Overexpression Lysate

N-Acetylmannosamine Kinase/GNE Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC222626

Accession#: NM_005476

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

79.1 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 N-Acetylmannosamine Kinase/GNE Overexpression Lysate

Western Blot: N-Acetylmannosamine Kinase/GNE Overexpression Lysate [NBP2-07744]

Western Blot: N-Acetylmannosamine Kinase/GNE Overexpression Lysate [NBP2-07744]

Western Blot: GNE Overexpression Lysate (Adult Normal) [NBP2-07744] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GNE.

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: N-Acetylmannosamine Kinase/GNE

The protein encoded by the GNE gene is a bifunctional enzyme that monitors and begins biosynthesis of N-acetylneuraminic acid (NeuAc). NeuAC is a precursor of sialic acids and functions in early development as sialylation is a component of cell adhesion, signal transduction, and tumorigencity and metastic behavior of malignant cells. The GNE gene has been linked to various diseases such as: myopathy, dementia, glomerulosclerosis, muscular dystrophy, and neuromuscular disease. This protein creates an enzyme that is rate-limiting in the sialic acid biosynthetic pathway as well as plays a role in amino and nucleotide sugar metabolism as well as CMP-N-acetylneuraminate biosynthesis in eukaryotes. The GNE gene is known to interact with genes CRMP1. ZBTB16, PIK3C2A, KIAA1549, and RIF1 to participate in these pathways. GNE exists is five isoforms: isoform 1: 722 amino acids long; 79 kDA; isoform 2: 753 amino acids long, 83 kDA; isoform 3: 681 amino acids long; 74 kDA; isoform 4: 648 amino acids long, 71 kDA; isoform 5: 612 amino acids long, 66 kDA.

Long Name

Glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine Kinase

Alternate Names

DMRV, GLCNE, GNE, IBM2, NAcetylmannosamine Kinase, Uae1

Gene Symbol

GNE

Additional N-Acetylmannosamine Kinase/GNE Products

Product Documents for N-Acetylmannosamine Kinase/GNE 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 N-Acetylmannosamine Kinase/GNE 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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