NDUFA9 Overexpression Lysate

Western Blot: NDUFA9 Overexpression Lysate (Adult Normal) [NBL1-13546] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NDUFA9.

• Reactivity: Hu
• Applications: WB

NDUFA9 Overexpression Lysate Summary

• Description:

  NDUFA9 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC203464
  
  Accession#: NM_005002
  
  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: NDUFA9

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: 42.3 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 NDUFA9 Overexpression Lysate

• CC6
• CI39k
• CI-39k
• CI-39kD
• complex I 39kDa subunit
• Complex I-39kD
• MGC111043
• NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 9 (39kD)
• NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 9, 39kDa
• NADH dehydrogenase (ubiquinone) Fe-S protein 2-like (NADH-coenzyme Q reductase)
• NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9, mitochondrial
• NADH-ubiquinone oxidoreductase 39 kDa subunit
• NDUFS2L
• SDR22E1
• short chain dehydrogenase/reductase family 22E, member 1

Background

ATP is generated via oxidative phosphorylation (Oxphos) in the mitochondria of almost all cells. The protein components of Oxphos, 4 respiratory chain complexes (I-IV) and an ATP synthase, are encoded in both the mitochondrial and nuclear genomes. Of the 4 respiratory chain complexes, complex I is the largest at 900 kD and contains at least 41 polypeptide subunits, 7 of which are encoded in the mitochondria, the remaining subunits are encoded in the nucleus. The multisubunit NADH:ubiquinone oxidoreductase is the first enzyme complex. By use of chaotropic agents, complex I can be fragmented into 3 different fractions. The flavoprotein fraction contains the NDUFV1, NDUFV2, and NDUFV3 subunits. The iron-sulfur protein (IP) fraction contains at least 7 subunits, NDUFS1-NDUFS6 and NDUFA5. The remaining subunits are part of the hydrophobic protein (HP) fraction. NDUFA9 is part of the hydrophobic protein fraction of the enzyme complex. However, it is predominantly hydrophilic, and appears to lie mostly outside the lipid bilayer.

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: NDUFA9