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PrecisION hKv7.2/hKv7.3-CHO Recombinant Cell Line
Description:
PrecisION hKv7.2/hKv7.3-CHO Recombinant Cell Line
Trade Name:
Upstate (Millipore)
Qty/Pk:
2 mL
Product Overview:
Recombinant CHO-K1 cell line co-expressing the human Kv7.2 [voltage-gated potassium channel, KQT-like subfamily, member 2 (KCNQ2), accession number Y15065] and the human Kv7.3 [voltage-gated potassium channel, KQT-like subfamily, member 3 (KCNQ3), accession number NM_004519].
Format:
2 x 1 ml aliquots containing 1.58 x 106cells/ml in 7.5% DMSO at passage 10.
Format:
2 x 1 ml aliquots containing 1.58 x 106cells/ml in 7.5% DMSO at passage 10.
Background Information:
Kv7.2 and Kv7.3 are both voltage-gated, delayed rectifier potassium channels. Together they co assemble to form the M current in the brain. The M current is a slowly activating and deactivating potassium conductance that plays a crucial role in determining the excitability of neurons as well as their responsiveness to synaptic inputs. This current is also expressed in many neurons in the central nervous system. Enhancers of the M current are used for antiepileptic therapy and blockers have been shown to enhance learning and memory.
Application Notes:
hKv7.2/hKv7.3 (KCNQ2/KCNQ3) channels have been selectively expressed in a CHO-K1 cell line and characterized in terms of their biophysical and pharmacological properties using whole-cell and perforated patch clamp techniques and IonWorksTM Quattro. The currents displayed all the hallmarks for selective expression of Kv7.x channels such as a relatively negative threshold of activation, the absence of any inactivation and slow activation/deactivation kinetics. The current/voltage relationship was linear and the mean outward current at 0 mV was 2.2 ± 0.3 nA (n=3). Thus this cell line has an ideal level of functional expression for both manual and automated patch clamp applications. Using perforated patch clamp techniques the currents were found to be dose-dependently blocked by TEA in the appropriate range for expression of Kv7.2/Kv7.3 channels (approximately 58% inhibition at 3 mM). The effects of the selective Kv7.x blockers linopirdine and XE991 were also assessed using IonWorksTM Quattro and the IC50 values obtained were similar to published values; around 9 μM and 2 μM respectively. Channel expression, monitored using IonWorksTM HT, is robust over at least 40 passages. For example 96% of cells expressed outward current greater than 500 pA at passage 40 (n=170) with a mean current amplitude of 2 +/-0.06 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.
Entrez Gene Summary:
The M channel is a slowly activating and deactivating potassium channel that plays a critical role in the regulation of neuronal excitability. The M channel is formed by the association of the protein encoded by this gene and a related protein encoded by the KCNQ3 gene, both integral membrane proteins. M channel currents are inhibited by M1 muscarinic acetylcholine receptors and activated by retigabine, a novel anti-convulsant drug. Defects in this gene are a cause of benign familial neonatal convulsions type 1 (BFNC), also known as epilepsy, benign neonatal type 1 (EBN1). At least five transcript variants encoding five different isoforms have been found for this gene.
UniProt Summary:
FUNCTION: SwissProt: O43526 # Probably important in the regulation of neuronal excitability. Associates with KCNQ3 to form a potassium channel with essentially identical properties to the channel underlying the native M-current, a slowly activating and deactivating potassium conductance which plays a critical role in determining the subthreshold electrical excitability of neurons as well as the responsiveness to synaptic inputs. KCNQ2/KCNQ3 current is blocked by linopirdine and XE991, and activated by the anticonvulsant retigabine. Muscarinic agonist oxotremorine-M strongly suppress KCNQ2/KCNQ3 current in cells in which cloned KCNQ2/KCNQ3 channels were coexpressed with M1 muscarinic receptors.
SIZE: 872 amino acids; 95848 Da
SUBUNIT: Heteromultimer with KCNQ3. May associate with KCNE2.
SUBCELLULAR LOCATION: Membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: In adult and fetal brain. Highly expressed in areas containing neuronal cell bodies, low in spinal chord and corpus callosum. Isoform 2 is preferentially expressed in differentiated neurons, whereas isoform 6 is prominent in fetal brain, undifferentiated neuroblastoma cells, and brain tumors.
DOMAIN: SwissProt: O43526 The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position (By similarity).
PTM: In Xenopus oocytes KCNQ2/KCNQ3 heteromeric current can be increased by intracellular cyclic AMP, an effect that depends on phosphorylation of Ser-52 in the N-terminus region.
DISEASE: SwissProt: O43526 # Defects in KCNQ2 are the cause of benign familial neonatal convulsions type 1 (BFNC1) [MIM:121200]; also known as epilepsy, benign neonatal type 1 (EBN1). BFNC1 is an autosomal dominant form of epilepsy in the newborn that clears spontaneously after a few weeks and is followed by normal psychomotor development. & Defects in KCNQ2 are the cause of myokymia with neonatal epilepsy [MIM:606437]. Neonatal epilepsy or benign familial neonatal convulsions (BFNC) is a generalized, idiopathic epilepsy syndrome of the newborn. Neonatal epilepsy with myokymia consists of a syndrome in which neonatal epilepsy is followed later in life by myokymia, the spontaneous involuntary contractions of skeletal muscles fiber groups that can be observed as vermiform movement of the overlying skin. Electromyography typically shows continuous motor unit activity with spontaneous oligo- and multiplet- discharges of high intraburst frequency (myokymic discharges).
SIMILARITY: Belongs to the potassium channel family. KQT subfamily.
MISCELLANEOUS: Inclusion of isoform 6 in heteromultimers results in attenuation of potassium current. Prominent expression of isoform 6 in the developing brain may alter firing repertoires of immature neurons excitability to provide cues for proliferation rather than differentiation. & Mutagenesis experiments were carried out in Xenopus oocytes by coexpression of either KCNQ2(mut) and KCNQ3 at the ratio of 1:1, or of KCNQ2(mut), KCNQ2(wt) and KCNQ3 at the ratio of 1:1:2, to mimic the situation in a heterozygous patient with BFNC1 disease.
SIZE: 872 amino acids; 95848 Da
SUBUNIT: Heteromultimer with KCNQ3. May associate with KCNE2.
SUBCELLULAR LOCATION: Membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: In adult and fetal brain. Highly expressed in areas containing neuronal cell bodies, low in spinal chord and corpus callosum. Isoform 2 is preferentially expressed in differentiated neurons, whereas isoform 6 is prominent in fetal brain, undifferentiated neuroblastoma cells, and brain tumors.
DOMAIN: SwissProt: O43526 The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position (By similarity).
PTM: In Xenopus oocytes KCNQ2/KCNQ3 heteromeric current can be increased by intracellular cyclic AMP, an effect that depends on phosphorylation of Ser-52 in the N-terminus region.
DISEASE: SwissProt: O43526 # Defects in KCNQ2 are the cause of benign familial neonatal convulsions type 1 (BFNC1) [MIM:121200]; also known as epilepsy, benign neonatal type 1 (EBN1). BFNC1 is an autosomal dominant form of epilepsy in the newborn that clears spontaneously after a few weeks and is followed by normal psychomotor development. & Defects in KCNQ2 are the cause of myokymia with neonatal epilepsy [MIM:606437]. Neonatal epilepsy or benign familial neonatal convulsions (BFNC) is a generalized, idiopathic epilepsy syndrome of the newborn. Neonatal epilepsy with myokymia consists of a syndrome in which neonatal epilepsy is followed later in life by myokymia, the spontaneous involuntary contractions of skeletal muscles fiber groups that can be observed as vermiform movement of the overlying skin. Electromyography typically shows continuous motor unit activity with spontaneous oligo- and multiplet- discharges of high intraburst frequency (myokymic discharges).
SIMILARITY: Belongs to the potassium channel family. KQT subfamily.
MISCELLANEOUS: Inclusion of isoform 6 in heteromultimers results in attenuation of potassium current. Prominent expression of isoform 6 in the developing brain may alter firing repertoires of immature neurons excitability to provide cues for proliferation rather than differentiation. & Mutagenesis experiments were carried out in Xenopus oocytes by coexpression of either KCNQ2(mut) and KCNQ3 at the ratio of 1:1, or of KCNQ2(mut), KCNQ2(wt) and KCNQ3 at the ratio of 1:1:2, to mimic the situation in a heterozygous patient with BFNC1 disease.
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:
CHO
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:
Chinese Hamster Ovarian K-1 cells (CHO K-1)
Presentation:
2 x 1 ml aliquots
Protein Target:
hKv7.2/hKv7.3
UniProt Number:
Target Sub-Family:
Potassium
Packaging:
2 x 1 ml aliquots
Gene Symbol:
- O43526
- KVEBN1
- EBN
- KV7.2
- ENB1
- KCNA11
- HNSPC
- Kv7.2
- EBN1
- BFNC
Protein or Enzyme Type:
Ion Channels
Product Name:
PrecisION hKv7.2/hKv7.3-CHO Recombinant Cell Line
Entrez Gene Number:
Alternate Names:
- O43526
- KVEBN1
- EBN
- KV7.2
- ENB1
- KCNA11
- HNSPC
- Kv7.2
- EBN1
- BFNC