PEA-15 Overexpression Lysate

Western Blot: PEA-15 Overexpression Lysate [NBL1-14273] - Western blot validation of overexpression lysate (Cat # NBL1-14273) using anti-DDK antibody (Cat # NBP1-71705). Left: Protein ladder; Right: Cell lysates from HEK293T cells transfected with PEA-15 Human Tagged ORF Clone using transfection reagent PEI.

• Reactivity: Hu
• Applications: WB

PEA-15 Overexpression Lysate Summary

• Description:

  PEA-15 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC205507
  
  Accession#: NM_003768
  
  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: PEA15

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: 14.9 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 PEA-15 Overexpression Lysate

• HMAT1
• HMAT115kD
• HUMMAT1H
• MAT1H
• PEA15
• PEA-15
• PED
• PED15 kDa phosphoprotein enriched in astrocytes
• phosphoprotein enriched in astrocytes 15
• Phosphoprotein enriched in diabetes

Background

PEA-15 (phosphoprotein enriched in astrocytes) exists in an non-phosphorylated form (N), and two phosphorylated forms, Pa and Pb. PEA-15 is an endogenous substrate for PKC, which mediates the transition from Pa to Pb. The level of PEA-15 phosphorylation changes upon depolymerization or stabilization of tubulins, indicating that PEA-15 co-localizes with microtubules. The first 80 amino acids of PEA-15 correspond to the death effector domain (DED), which is a domain found in proteins that regulate apoptotic signaling pathways. The DED domain is necessary for PEA-15 to block Ras suppression. Although PEA-15 is predominantly expressed in the central nervous system, low levels of PEA-15 are expressed in liver and kidney, and higher levels in muscle. PEA-15 is also referred to as PED, phosphoprotein enriched in diabetes, for its elevated expression in type 2 diabetic patients.

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: PEA15