PDE7A Overexpression Lysate

Western Blot: PDE7A Overexpression Lysate (Adult Normal) [NBP2-08091] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PDE7A.

• Reactivity: Hu
• Applications: WB

PDE7A Overexpression Lysate Summary

• Description:

  PDE7A Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC220940
  
  Accession#: NM_002604
  
  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: PDE7A

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: 48.6 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 PDE7A Overexpression Lysate

• EC 3.1.4
• EC 3.1.4.17
• HCP1high affinity cAMP-specific 3'-5'-cyclic phosphodiesterase 7A
• PDE7
• PDE7A
• phosphodiesterase 7A
• phosphodiesterase isozyme 7
• TM22

Background

The cyclic monophosphate nucleotides (cyclic adenosine monophosphate [cAMP] and cyclic guanosine monophosphate [cGMP]) are found ubiquitously in mammalian cells and act as second messenger transducers to effect the intracellular actions of a variety of G protein coupled receptors (GPCRs) for hormones, cytokines, and neurotransmitters. Cyclic nucleotides are important intracellular second messengers which play important role in a variety of signal transduction process. The cyclic nucleotides are hydrolyzed and compartmentalized by a family of enzymes called phosphodieterases. One of the many phosphodiesterases that compartmentalized and hydrolyze cAMP in T Lymphocytes are phosphodiesterase type 7. The cAMP specific phosphodiesterase type 7 (PDE7) family is comprised of 2 genes (PDE7A and PDE7B) each with multiple splice variants generated by RNA splicing and use of alternate initiation sites. PDE7A has two splice variants (PDE7A1 and PDE7A2). The two are divergent at the 5 prime end in human, and PDE7A1 is more hydrophobic. Like other PDEs, human PDE7A gene has 456 amino acids and migrates at an apparent molecular weight of 53 to 55 kDa on reduced SDS PAGE. The PDE7A has a significant conserved region of about 270 amino acids common to all PDEs at the carboxy terminal apparently serves as the catalytic domain. The amino terminal region of this protein is divergent and presumably accounts for the distinctive and regulatory properties unique to the individual PDE families. PDE7A protein showed significant homology to other cAMP dependent PDEs (23%) with in the catalytic domain. PDE7A is widely expressed in various tissues including skeletal muscle, T lymphocytes, brain and pancreas.

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: PDE7A