PP1 Inhibitor-2 Overexpression Lysate

Western Blot: Protein phosphatase 1 inhibitor subunit 2 Overexpression Lysate (Adult Normal) [NBL1-14682] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Protein phosphatase 1 inhibitor subunit 2.

• Reactivity: Hu
• Applications: WB

PP1 Inhibitor-2 Overexpression Lysate Summary

• Description:

  PP1 Inhibitor-2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200280
  
  Accession#: NM_006241
  
  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: PPP1R2

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: 22.8 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 PP1 Inhibitor-2 Overexpression Lysate

• IPP2
• IPP-2
• IPP2phosphatase inhibitor 2
• MGC87148
• phosphoprotein phosphatase
• PP1 Inhibitor2
• PP1 Inhibitor-2
• PPP1R2
• protein phosphatase 1, regulatory (inhibitor) subunit 2
• protein phosphatase inhibitor 2

Background

Two inhibitors of protein phosphatase 1 (PP1) include the phosphatase inhibitor 1 (IPP-1) and phosphatase inhibitor 2 (IPP-2). IPP-2, also known as I-2, interacts with the catalytic subunit of PP1 to form the heterodimer PP1I. The PP1I complex is present in the cytosol of cells in a broad range of vertebrate and invertebrate species. Although the heterodimer itself is inactive, a reversible phosphorylation of IPP-2 at Thr 72 by glycogen-synthase-kinase (GSK3) initiates activation of the heterodimer complex in vitro. Phosphorylation of IPP-2 by casein kinase-II at Ser 86, Ser 120, and Ser 121 enhances the rate of phosphorylation by GSK3 at Thr 72 and effectively activates the heterodimer complex. Besides moderating PP1 activity, IPP-2 may play a role as a chaperone for the correct folding of PP1. The gene for human IPP-2 maps to chromosome 6 in the major histocompatiblity complex region.

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: PPP1R2