KCNK10 Overexpression Lysate

Western Blot: KCNK10 Overexpression Lysate (Adult Normal) [NBP2-06556] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KCNK10.

Product Discontinued

KCNK10 Overexpression Lysate Summary

• Description: Transient overexpression lysate of potassium channel, subfamily K, member 10 (KCNK10), transcript variant 1 The lysate was created in HEK293T cells, using Plasmid ID RC222775 and based on accession number NM_021161. The protein contains a C-MYC/DDK Tag. 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: KCNK10

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: 59.6 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

The lysate was created in HEK293T cells, using plasmid ID RC222775 and based on accession number NP_066984. The protein contains a C-terminal DDK tag. 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.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for KCNK10 Overexpression Lysate

• 2P domain potassium channel TREK2
• K2p10.1
• KCNK10
• Outward rectifying potassium channel protein TREK-2
• potassium channel subfamily K member 10
• potassium channel TREK-2
• potassium channel, subfamily K, member 10
• TREK-2 K(+) channel subunit
• TREK2
• TREK-2
• TREK2FLJ43399
• TWIK-related K+ channel 2

Background

KCNK10, also known as Potassium channel subfamily K member 10, has 3 isoforms, Isoform A that is 538 amino acid and app. 60 kDa, Isoform B that is 543 amino acid and app. 60 kDa, and Isoform C That is 543 amino acid and app. 60 kDa; expressed in pancreas and kidney and to a lower level in brain, testis, colon, and small intestine; belongs to the family of potassium channel proteins containing two pore-forming P domains; outwards rectifying potassium channel, produces rapidly activating and non-inactivating outward rectifier K(+) currents, activated by arachidonic acid and other naturally occurring unsaturated free fatty acids. Studies are being performed on the relationship of this protein with dentin sensitivity, brain ischemia, and ischemia. The protein has been shown to interact with AKAP150 protein and has been involved in several pathways including cholera infection, neuropathic pain-signaling in dorsal horn neurons, hepatic ABC transporters, PKA signaling, potassium transporters - outward current, gastric, and sweet taste signaling.

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: KCNK10