O-GlcNAc Transferase p110 subunit Overexpression Lysate

Western Blot: O-GlcNAc Transferase p110 subunit Overexpression Lysate (Adult Normal) [NBP2-04900] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for O-GlcNAc Transferase p110 subunit.

• Reactivity: Hu
• Applications: WB

O-GlcNAc Transferase p110 subunit Overexpression Lysate Summary

• Description:

  O-GlcNAc Transferase p110 subunit Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC224481
  
  Accession#: NM_181673
  
  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: OGT

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: 115.5 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.

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 O-GlcNAc Transferase p110 subunit Overexpression Lysate

• EC 2.4.1
• EC 2.4.1.255
• FLJ23071
• HRNT1
• MGC22921
• O-GlcNAc transferase p110 subunit
• O-GlcNAc transferase subunit p110
• OGlcNAc Transferase
• O-GlcNAc Transferase
• O-GLCNAC
• OGT
• O-linked N-acetylglucosamine (GlcNAc) transferase(UDP-N-acetylglucosamine:polypeptide-N-acetylglucosaminyl transferase)
• O-linked N-acetylglucosamine transferase 110 kDa subunit
• UDP-N-acetylglucosamine--peptide N-acetylglucosaminyltransferase 110 kDasubunit
• uridinediphospho-N-acetylglucosamine:polypeptide beta-N-acetylglucosaminyltransferase

Background

Diffusion of metabolites and small non-nuclear molecules as well as active, mediated import of protein and export of protein and RNA through the nuclear envelope occurs through nuclear pore complexes or NPC's. NPC's contain up to 100 different polypeptides which have a combined mass of about 125 megadaltons. The channel available for passive transport through the NPC is about 9-10 nm in diameter while carrier mediated changes in the NPC result in a ~25 nm channel used for larger, actively transported molecules. Of the 100 polypeptides, at least 8 of these are O-linked N-acetylglycosamine-modified in mammalian cells. All of the mammalian O-linked glycoproteins contain multiple copies of phenylalanine, glycine dipeptide repeats dispersed throughout part of their sequence. Studies indicate that the NPC O-linked glycoproteins have a direct role in nuclear protein import.

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: OGT