IFIT2 Overexpression Lysate

Western Blot: IFIT2 Overexpression Lysate (Adult Normal) [NBL1-11835] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IFIT2.

• Reactivity: Hu
• Applications: WB

IFIT2 Overexpression Lysate Summary

• Description:

  IFIT2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC205582
  
  Accession#: NM_001547
  
  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: IFIT2

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: 54.5 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 IFIT2 Overexpression Lysate

• cig42
• G10P2IFI-54K
• GARG-39
• IFI-54
• IFI54IFIT-2
• IFI-54K
• Interferon, alpha-inducible protein (MW 54kD)
• Interferon-induced 54 kDa protein
• interferon-induced protein 54
• interferon-induced protein with tetratricopeptide repeats 2
• ISG-54 K
• ISG54
• ISG-54K
• P54

Background

Saha et al. used this antibody to study protein expression of murine GARG39/IFIT2 in relation to viral infection. Immunofluorescence localized this protein in association with the mitotic spindle in both mitotically active normal NIH3T3 and B16F10 melanoma cells. Western blotting shows approx. 55 kD band GARG39/IFIT2 in brain tissue of mice infected with Japanese encephalitis virus (JEV). Brain tissue from non-infected mice did not display this band. In 1994 Bluyssen et al. characterized the gene for murine GARG39/IFIT2 and historically the protein has been identified as interferon-induced protein with tetratricopeptide repeats 2 (IFIT-2), interferon-induced 54 kDa protein (IFI-54K), glucocorticoid-attenuated response gene 39 protein (GARG-39). Human IFIT2, with 62% amino acid sequence identity, has been referred to as Interferon-induced protein with tetratricopeptide repeats 2 (IFIT-2), Interferon-induced 54 kDa protein (IFI-54K) and ISG-54 K. Kang et al. showed with proteome analysis IFIT2 was expressed more than two-fold in human osteosarcoma cells U2OS treated with ascochlorin. Kawada et al. studied gene expression to show that IFIT2 is strongly upregulated in peripheral blood in children during the acute phase of influenza.

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: IFIT2