N-Acetylmannosamine Kinase/GNE Overexpression Lysate

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.

• Reactivity: Hu
• Applications: WB

N-Acetylmannosamine Kinase/GNE Overexpression Lysate Summary

• Description:

  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.
• Gene: GNE

Applications/Dilutions

• Dilutions:
  • Western Blot
• 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: 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.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

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.

Alternate Names for N-Acetylmannosamine Kinase/GNE Overexpression Lysate

• bifunctional UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase
• DMRV
• GLCNE
• GLCNENM
• glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine kinase
• GNE
• IBM2
• NAcetylmannosamine Kinase
• N-Acetylmannosamine Kinase
• N-acylmannosamine kinase
• Uae1
• UDP-GlcNAc-2-epimerase/ManAc kinase
• UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase
• UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase

Background

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.

Limitations

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.

Bioinformatics

• Gene Symbol: GNE