PUS10 Overexpression Lysate

Western Blot: PUS10 Overexpression Lysate (Adult Normal) [NBL1-14998] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PUS10.

• Reactivity: Hu
• Applications: WB

PUS10 Overexpression Lysate Summary

• Description:

  PUS10 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210979
  
  Accession#: NM_144709
  
  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: PUS10

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: 60.1 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 PUS10 Overexpression Lysate

• CCDC139
• coiled-coil domain containing 139
• Coiled-coil domain-containing protein 139
• DOBIMGC126729
• EC 5.4.99
• EC 5.4.99.-
• FLJ32312
• MGC126755
• pseudouridine synthase 10
• pseudouridylate synthase 10
• Psi55 synthase
• PUS1
• putative tRNA pseudouridine synthase Pus10
• tRNA pseudouridine 55 synthase
• tRNA pseudouridylate synthase
• tRNA-uridine isomerase

Background

Pseudouridination, the isomerization of uridine to pseudouridine, is the most common posttranscriptional nucleotidemodification found in RNA and is essential for biologic functions such as spliceosome biogenesis. Pseudouridylatesynthases, such as PUS10, catalyze pseudouridination of structural RNAs, including transfer, ribosomal, and splicingRNAs. These enzymes also act as RNA chaperones, facilitating the correct folding and assembly of tRNAs (McCleverty etal., 2007 (PubMed 17900615)).(supplied by OMIM)

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: PUS10