TIMM50 Overexpression Lysate

Western Blot: TIMM50 Overexpression Lysate (Adult Normal) [NBP2-08987] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TIMM50.

• Reactivity: Hu
• Applications: WB

TIMM50 Overexpression Lysate Summary

• Description:

  TIMM50 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC224744
  
  Accession#: NM_001001563
  
  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: TIMM50

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: 50.3 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 TIMM50 Overexpression Lysate

• homolog of yeast Tim50
• MGC102733
• mitochondrial import inner membrane translocase subunit TIM50
• TIM50
• TIM50L
• Tim50-like protein
• translocase of inner mitochondrial membrane 50 homolog (S. cerevisiae)
• translocase of inner mitochondrial membrane 50 homolog (yeast)

Background

Tim50 is a component of the yeast Tim23 import machinery, which mediates translocation of presequence containing proteins across the mitochondrial inner membrane. By searching sequence databases, open reading frames encoding proteins with similarity to Tim50 and with a putative mitochondrial presequence were identified in the genomes of evolutionarily distant organisms, including human. Tim50 was found to interact with the N terminal intermembrane space domain of Tim23. Functional defects of Tim50 either by depletion of the protein or addition of anti Tim50 antibodies blocked the protein translocation across the inner membrane. A translocation intermediate accumulated at the translocator of the outer mitochondrial membrane (TOM) complex was cross linked to Tim50. Thus it can be concluded that Tim50, in cooperation with Tim23, facilitates transfer of the translocating protein from the TOM complex to the Tim23 complex.

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: TIMM50