PDE4B Overexpression Lysate

Western Blot: PDE4B Overexpression Lysate (Adult Normal) [NBP2-08546] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PDE4B.

• Reactivity: Hu
• Applications: WB

PDE4B Overexpression Lysate Summary

• Description:

  PDE4B Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210912
  
  Accession#: NM_001037341
  
  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: PDE4B

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: 83.2 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 PDE4B Overexpression Lysate

• cAMP-specific (phosphodiesterase E4 dunce homolog
• cAMP-specific 3'-5'-cyclic phosphodiesterase 4B
• cAMP-specific phosphodiesterase-4 B isoform
• DKFZp686F2182
• DPDE4
• Drosophila)
• dunce-like phosphodiesterase E4
• EC 3.1.4
• EC 3.1.4.17
• MGC126529
• PDE32
• PDE4B
• PDEIVB
• phosphodiesterase 4B, cAMP-specific (dunce (Drosophila)-homologphosphodiesterase E4)
• phosphodiesterase 4B, cAMP-specific (dunce
• phosphodiesterase 4B, cAMP-specific
• phosphodiesterase E4 dunce homolog

Background

PDE4B is a member of the type IV, cyclic AMP (cAMP)-specific, cyclic nucleotide phosphodiesterase (PDE) family. Enzymes of the cAMP-dependent phosphodiesterase type 4 (PDE4) family are important in hydrolyzing cAMP produced by G-protein coupled receptor (GPCR) stimulated adenylyl cyclases. Four genes (PDE4A, B, C and D) make up the PDE4 family. cAMP-dependent phosphodiesterase type B (PDE4B) family consists of four known splice variants, PDE4B1, B2, B3 and a novel 66 kDa PDE4B4 protein. One or more subtype variants of PDE4B are ubiquitously present in all mammalian cells. In CNS, all four PDE4B subtype variants are expressed in varying ratios and their activity is regulated in tandem with GPCRs stimulation. Some PDE4B variants are also substrate for PKA-dependent phosphorylation. Peripheral tissues exhibit differential expression of PDE4B1, B2 and B4 variants; the PDE4B4 (66kDa protein) is the major PDE4B variant expressed in lungs, testis and heart. In testis PDE4B enzymes are expressed in somatic cells (Sertoli and Leydig cells) while PDE4A is localized in germ cells.

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: PDE4B