epithelial Sodium Channel alpha Overexpression Lysate

Western Blot: epithelial Sodium Channel alpha Overexpression Lysate (Adult Normal) [NBP2-10616] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for epithelial Sodium Channel alpha.

• Reactivity: Hu
• Applications: WB

epithelial Sodium Channel alpha Overexpression Lysate Summary

• Description:

  epithelial Sodium Channel alpha Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC209564
  
  Accession#: NM_001038
  
  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: SCNN1A

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: 75.5 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 epithelial Sodium Channel alpha Overexpression Lysate

• alpha ENaC-2
• Alpha-ENaC
• Alpha-NaCH
• amiloride-sensitive epithelial sodium channel alpha subunit
• amiloride-sensitive sodium channel subunit alpha 2
• amiloride-sensitive sodium channel subunit alpha
• BESC2
• ENaCA
• ENaCalpha
• Epithelial Na(+) channel subunit alpha
• epithelial Sodium Channel alpha
• FLJ21883
• nasal epithelial sodium channel alpha subunit
• Nonvoltage-gated sodium channel 1 subunit alpha
• SCNEA
• SCNN1
• SCNN1A
• sodium channel, nonvoltage-gated 1 alpha

Background

Epithelial Sodium Channel alpha (SCNN1A) is a subunit of the epithelial sodium channel (ENaC). ENac has high sodium selectivity, low conductance, and amiloride sensitivity. The functional channel of ENaC is composed of at least 3 subunits, alpha (SCNN1A), beta (SCNN1B), and gamma (SCNN1G). The 3 subunits show sequence similarities to one another, indicating descent from a common ancestral gene. Each encodes a protein containing 2 transmembrane domains, with intracellular amino and carboxyl termini. The alpha subunit supports sodium conductance when expressed alone; the beta and gamma subunits do not support sodium conductance by themselves, but greatly augment the channel activity when expressed in conjunction with the alpha subunit. ENaC in the kidney, lung and colon plays an essential role in trans- epithelial sodium and fluid balance. ENaC also mediates aldosterone-dependent sodium re-uptake in the distal nephron of the kidney, thus regulating blood pressure. Gain-of-function mutations in beta- or gamma-ENaC can cause severe arterial hypertension (Liddel's syndrome) and loss-of-function mutations in alpha- or beta-ENaC causes pseudohypoaldosteronism (PHA-1).

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: SCNN1A