PDE8A Overexpression Lysate

Western Blot: PDE8A Overexpression Lysate (Adult Normal) [NBL1-14232] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PDE8A.

• Reactivity: Hu
• Applications: WB

PDE8A Overexpression Lysate Summary

• Description:

  PDE8A Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC209169
  
  Accession#: NM_002605
  
  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: PDE8A

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: 93.1 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 PDE8A Overexpression Lysate

• cAMP-specific cyclic nucleotide phosphodiesterase 8A
• EC 3.1.4.17
• FLJ16150
• high affinity cAMP-specific and IBMX-insensitive 3'-5'-cyclic phosphodiesterase8A
• high-affinity cAMP-specific and IBMX-insensitive 3'-5'-cyclic phosphodiesterase8A
• HsT19550
• PDE8A
• phosphodiesterase 8A

Background

Phosphodiesterases (PDEs) regulate the intracellular levels of cAMP and cGMP. These cyclic nucleotides play an important role as second messengers in multiple physiologic processes, including regulation of vascular resistance, cardiac output, visceral motility, immune response, inflammation, neuroplasticity, vision, and reproduction. PDEs comprise a large superfamily of enzymes divided into 10 families. Different PDEs can be distinguished by their structure, tissue expression, localization, substrate specificity, regulation, and sensitivity to PDE inhibitors. Diversity in structure and specificity of function make PDEs promising targets for the pharmacotherapy of diseases modulated by cyclic nucleotide signaling (Hetman et al., MIM 2000). See MIM 171885.[supplied by OMIM].

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: PDE8A