IRS2 Overexpression Lysate

Western Blot: IRS2 Overexpression Lysate (Adult Normal) [NBP2-07957] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IRS2.

• Reactivity: Hu
• Applications: WB

IRS2 Overexpression Lysate Summary

• Description:

  IRS2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC212413
  
  Accession#: NM_003749
  
  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: IRS2

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: 137.2 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 IRS2 Overexpression Lysate

• insulin receptor substrate 2
• IRS2
• IRS-2

Background

Insulin receptor substrates (IRS), the major intracellular substrates of the insulin receptor (IR), are adaptor proteins that transduce signals from the IR to downstream effectors that are important for the biological effect of insulin (1-2). After insulin stimulation, IRS proteins are rapidly phosphorylated on multiple tyrosine residues. Once phosphorylated, IRS proteins bind and activate Grb-2, SHP2 and the PI3-K p85 subunit (2-3). Sequences of IRS-2 and IRS-1 reveal a highly conserved amino terminus containing a pleckstrin-homology domain and a phosphotyrosine-binding domain, and a poorly conserved carboxy terminus containing several tyrosine phosphorylation motifs. IRS-2 is expressed in many cells, including tissues from IRS-1-/- mice, and may be essential for signaling by several receptor systems such as insulin and cytokine (1).

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: IRS2