eIF4A1 Overexpression Lysate

Western Blot: eIF4A1 Overexpression Lysate (Adult Normal) [NBL1-10201] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for eIF4A1.

• Reactivity: Hu
• Applications: WB

eIF4A1 Overexpression Lysate Summary

• Description:

  eIF4A1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC203298
  
  Accession#: NM_001416
  
  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: EIF4A1

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: 46 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 eIF4A1 Overexpression Lysate

• ATP-dependent RNA helicase eIF4A-1
• DDX2Aeukaryotic initiation factor 4A-I
• EC 3.6.1
• EC 3.6.4.13
• EIF4A
• EIF-4A
• eIF-4A-I
• eIF4A-I
• eukaryotic initiation factor 4AI
• eukaryotic translation initiation factor 4A
• eukaryotic translation initiation factor 4A, isoform 1
• eukaryotic translation initiation factor 4A1

Background

ATP-dependent RNA helicase which is a subunit of the eIF4F complex involved in cap recognition and isrequired for mRNA binding to ribosome. In the current model of translation initiation, eIF4A unwinds RNA secondarystructures in the 5'-UTR of mRNAs which is necessary to allow efficient binding of the small ribosomal subunit, andsubsequent scanning for the initiator codon

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: EIF4A1