KCNJ10 Overexpression Lysate

Western Blot: KCNJ10 Overexpression Lysate (Adult Normal) [NBP2-08133] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KCNJ10.

• Reactivity: Hu
• Applications: WB

KCNJ10 Overexpression Lysate Summary

• Description:

  KCNJ10 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC213543
  
  Accession#: NM_002241
  
  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: KCNJ10

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: 42.3 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 KCNJ10 Overexpression Lysate

• ATP-dependent inwardly rectifying potassium channel Kir4.1
• ATP-sensitive inward rectifier potassium channel 10
• BIRK-10
• glial ATP-dependent inwardly rectifying potassium channel KIR4.1
• Inward rectifier K(+) channel Kir1.2
• inward rectifier K+ channel KIR1.2
• KCNJ10
• KCNJ13-PEN
• KIR1.2
• KIR4.1
• Potassium channel, inwardly rectifying subfamily J member 10
• potassium inwardly-rectifying channel, subfamily J, member 10
• SESAME

Background

KCNJ10 encodes a member of the inward rectifier-type potassium channel family, characterized by having a greater tendency to allow potassium to flow into, rather than out of, a cell. The encoded protein may form a heterodimer with another potassium channel protein and may be responsible for the potassium buffering action of glial cells in the brain. Mutations in this gene have been associated with seizure susceptibility of common idiopathic generalized epilepsy syndromes.

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.

FAQs for KCNJ10 Lysate (NBP2-08133). (Showing 1 - 3 of 3 FAQ).

1. Does this product correspond to the full length protein? Can we detect kir4.1 in this lysate with specific Kir4.1 antibodies?
   • NBP2-08133 lysate was created in HEK293T cells, using plasmid ID RC215316 and based on accession number NP_002232 which corresponds to the full length KCNJ10 and this protein contains a C-terminal DDK tag. Technically speaking you should be able to use a Kir 4.1 antibody to detect Kir 4.1 in this lysate, however please note that our western blot image was generated using anti Flag antibody.
2. I would like to know which specific peptide sequence are detected by the antibody? It is only stated in the data sheet that it reacts against a part of the C-terminal part of KIR4.1. Do you know if this antibody or any of your other antibodies could be utilized as an intracellular staining of cells for flow cytometry? Furthermore, I have seen in the data sheet for the synthetic peptide that it is not suitable for use with live cells, but is this an absolute? If this is the case, would it possible to buy a similar peptide which could be used with live cells, and if so at which prize/amount? Lastly, are the peptide in a frozen liquid form (or solid powder) with out trace of azide? When it comes to the cell lysate I`m wondering if this could be utilized with live cells or if the RIPA buffer which it is dissolved in makes this very toxic? If the last is the case, is it possible to provide the lysate in another buffer which are appropriate for live cells? Lastly, I wondered if the transfected HEK293T cells are also sold for culture?
   • I think that the RIPA buffer would make this lysate unsuitable for live cells. This product is intended for use as a control in western blotting. NBP2-08133 is our only KCNJ10 lysate; furthermore, we do not sell HEK293T cells.
3. What is the volume of lysate provided?
   • The KCNJ10 overexpression lysate product NBP2-08133 includes one vial of overexpressed HEK293 cell lysate (100ug in 100ul), one vial of empty-vector transfected HEK293 cell lysate (100ug in 100ul), and a vial of 2X sample buffer.

Bioinformatics

• Gene Symbol: KCNJ10