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PrecisION hHCN2-HEK Recombinant Cell Line
Description:
PrecisION hHCN2-HEK Recombinant Cell Line
Trade Name:
Upstate (Millipore)
Qty/Pk:
2 mL
Product Overview:
Recombinant HEK293 cell line expressing the human hyperpolarization-activated, cyclic nucleotide-gated potassium channel 1 (HCN2).
Format:
2 x 1 ml aliquots containing 1.14 x 106cells/ml in 7.5% DMSO at passage 11.
Format:
2 x 1 ml aliquots containing 1.14 x 106cells/ml in 7.5% DMSO at passage 11.
Background Information:
Hyperpolarization-activated, cyclic nucleotide-modulated (HCN) channels contribute to rhythmic spontaneous electrical activity in the heart and CNS. Interference with HCN2 activity can lead to various forms of cardiac arrhythmias, due to dysfunction of the sinus node. The predominant role of HCN2 in these cells may be the stabilization of a normal diastolic membrane potential
Application Notes:
HEK293 cells expressing hHCN2 were characterised in terms of their biophysical and pharmacological properties using whole-cell patch clamp techniques and IonWorksTM HT electrophysiology. The biophysical properties are typical of hHCN2 both in terms of V½ of activation (-102.6 ± 0.3 mV, n=7) and the time course of activation (e.g. 4026 ± 712 ms at -100 mV and 670 ±140 ms at -130 mV, n=8). Under whole-cell recording conditions the mean current amplitude was 5.5 ± 1 nA at -130 mV (n =7) and reversal potential measurements indicated permeability to both Na+ and K+ (PNa/PK around 0.2) typical of HCN channels. Currents were dose-dependently inhibited by the selective HCN blocker ZD7288 (IC50 value of 25 µM) and the inorganic cation Caesium (Cs+, IC50 value of 119 µM) in line with reported literature values. The currents also exhibited a marked sensitivity to the tyrosine kinase inhibitor genistein (100 µM) inhibiting the current by approximately 66% in contrast to the effects of the same concentration on hHCN1 currents (greater than 10% inhibition, Millipore PrecisIONTM HCN1-HEK293 recombinant cell line, CYL3040). These differences in sensitivity have been noted by others and support the notion that this cell line selectively expresses hHCN2 channels. The currents were also sensitive to cAMP (100 µM) since intracellular application shifted the V½ of activation by +13 mV again typical of HCN2 channels. Channel expression, monitored using IonWorksTM HT, is robust over at least 40 passages. For example 68% of cells expressed outward current greater than 500 pA at passage 40 (n=117) with a mean current amplitude of 3.12 nA.
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Species:
Human
Quality Assurance:
Mycoplasma Testing:
The cell line has been screened to confirm the absence of Mycoplasma species
The cell line has been screened to confirm the absence of Mycoplasma species
Cell Type:
HEK
Cell Line Type:
Ion Channel Cell Lines
Components:
Pack contains 2 vials of mycoplasma-free cells, 1 ml per vial
Brand Family:
Upstate
Host Cells:
293 cell line
Protein Target:
hHCN2
Presentation:
2 x 1 ml aliquots
Target Sub-Family:
Hyperpolarized
Packaging:
2 x 1 ml aliquots
Protein or Enzyme Type:
Ion Channels
Product Name:
PrecisION hHCN2-HEK Recombinant Cell Line