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Kir2.2 Overexpression Lysate

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Western Blot: Kir2.2 Overexpression Lysate (Adult Normal) [NBP2-06575] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Kir2.2.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Kir2.2 Overexpression Lysate Summary

Description

Kir2.2 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC207051

Accession#: NM_021012

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
KCNJ12

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
48.8 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 Kir2.2 Overexpression Lysate

  • ATP-sensitive inward rectifier potassium channel 12
  • hIRK
  • hIRK1
  • hkir2.2x
  • Inward rectifier K(+) channel Kir2.2
  • Inward rectifier K(+) channel Kir2.2v
  • inward rectifier K(+) channel Kir2.6
  • IRK2
  • IRK-2
  • IRK2IRK-2
  • KCNJ12
  • kcnj12x
  • KCNJN1
  • KCNJN1FLJ14167
  • Kir2.2
  • Kir2.2v
  • Potassium channel, inwardly rectifying subfamily J member 12
  • potassium inwardly-rectifying channel, subfamily J, inhibitor 1
  • potassium inwardly-rectifying channel, subfamily J, member 12

Background

FUNCTION: Probably participates in establishing action potential waveform and excitability of neuronal and muscle tissues. Inward rectifier potassium channels are characterized by a greater tendency to allow potassium to flow into the cell rather than out of it. Their voltage dependence is regulated by the concentration of extracellular potassium; as external potassium is raised, the voltage range of the channel opening shifts to more positive voltages. The inward rectification is mainly due to the blockage of outward current by internal magnesium. Can be blocked by extracellular barium and cesium. Tissue specificity: Highest level in cerebellum. Moderately found in kidney, forebrain and skeletal muscle. Not detected in uterus, liver and pancreas. Subcellular location: Membrane, Multi-pass membrane protein.

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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Product General Protocols

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Video Protocols

WB Video Protocol

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Bioinformatics

Gene Symbol KCNJ12