ZNF474 Overexpression Lysate

Western Blot: ZNF474 Overexpression Lysate (Adult Normal) [NBP2-04510] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ZNF474.

• Reactivity: Hu
• Applications: WB

ZNF474 Overexpression Lysate Summary

• Description:

  ZNF474 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC222255
  
  Accession#: NM_207317
  
  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: ZNF474

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: 40.1 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 ZNF474 Overexpression Lysate

• 4933409D10Rik
• testis-specific zinc finger protein
• TSZFP
• zinc finger protein 474

Background

Chimera RNA interference (chimera RNAi) is process by which small interfering RNA/DNA chimera triggers the destruction of mRNA for the original gene. The discovery work, design, and application of chimera RNAi has been pioneered by Professor Kaoru Saigo and Dr. Kumiko Ui-Tei at the University of Tokyo. Chimera RNAi has many advantages over the conventional siRNAs. First, it has been demonstrated to have reliable knock-down for over 10,000 human genes. Because the human genome is composed of an intricate, genetic network, chimera RNAi's unique design has successfully obviated the off-target effects including microRNA-based influence. Another advantage of the chimera RNAi technology is its effectiveness at low concentrations (0.5nM to 5nM); only mRNA is destroyed so genomic genes are not affected. Finally, having both the sense and anti-sense strands consisting RNA/DNA chimera, it offers much greater compound stability for streamlining in vitro and in vivo assays and applications while minimizing interferon induction and other adverse reactions.

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: ZNF474