IRF8 Overexpression Lysate

Western Blot: IRF8 Overexpression Lysate (Adult Normal) [NBP2-08141] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IRF8.

• Reactivity: Hu
• Applications: WB

IRF8 Overexpression Lysate Summary

• Description:

  IRF8 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC217646
  
  Accession#: NM_002163
  
  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: IRF8

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: 48.2 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 IRF8 Overexpression Lysate

• ICSBP1
• ICSBPH-ICSBP
• interferon consensus sequence binding protein 1
• Interferon consensus sequence-binding protein
• interferon regulatory factor 8
• IRF8
• IRF-8ICSBP1

Background

Interferon regulatory factor 8 (IRF8) / Interferon consensus sequence binding protein (ICSBP) is a transcription factor of the interferon (IFN) regulatory factor (IRF) family. Proteins of this family are composed of a conserved DNA binding domain in the N terminal region and a divergent C terminal region that serves as the regulatory domain. The IRF family proteins bind to the IFN stimulated response element (ISRE) and regulate expression of genes stimulated by type I IFNs, namely IFN alpha and IFN beta. IRF family proteins also control expression of IFN alpha and IFN beta regulated genes that are induced by viral infection. IRF8 specifically binds to the upstream regulatory region of type I IFN and IFN inducible MHC class I genes (the interferon consensus sequence (ICS)). Unlike IRF3, IRF8 appears to act as a negative regulator of IFN induced genes in most cases, but IRF8 mediates activation of NF kappaB by the toll like receptor 9 (TLR9) after stimulation by unmethylated CpG DNA in dendritic cells. Finally, it has been shown that IRF8 decreases bcl-2 expression and thus may play a role in chronic myelogenous leukemia.

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: IRF8