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epithelial Sodium Channel beta Overexpression Lysate

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Western Blot: epithelial Sodium Channel beta Overexpression Lysate (Adult Normal) [NBL1-15746] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for epithelial Sodium ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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epithelial Sodium Channel beta Overexpression Lysate Summary

Description

epithelial Sodium Channel beta Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC206580

Accession#: NM_000336

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
SCNN1B

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
72.5 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 beta Overexpression Lysate

  • amiloride-sensitive sodium channel subunit beta 1
  • amiloride-sensitive sodium channel subunit beta
  • BESC1
  • Beta-ENaC
  • Beta-NaCH
  • ENaCB
  • ENaCbeta
  • Epithelial Na(+) channel subunit beta
  • epithelial Sodium Channel beta
  • epithelial sodium channel beta-2 subunit
  • epithelial sodium channel beta-3 subunit
  • nasal epithelial sodium channel beta subunit
  • Nonvoltage-gated sodium channel 1 subunit beta
  • SCNEB
  • SCNN1B
  • sodium channel, nonvoltage-gated 1, beta

Background

SCNN1B 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. 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. Mutations in beta or gamma subunit of the epithelial sodium channel (ENaC) have been found to cause a hereditary form of human hypertension, Liddle syndrome.

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.

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Bioinformatics

Gene Symbol SCNN1B