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PrecisION hTRPA1-HEK Recombinant Cell Line
Description:
PrecisION hTRPA1-HEK Recombinant Cell Line
Trade Name:
PrecisION
Qty/Pk:
2 mL
Product Overview:
Recombinant HEK293 cell line expressing the human TRPA1 ligand gated ion channel
Background Information:
There is now compelling evidence to suggest that TRPA1 is primarily responsible for mediating sensations of noxious cold (<17 deg) and has a significant role in conveying inflammatory pain responses (Garcia-Anoveros and Nagata, 2007). TRPA1 is located in a subset of DRG neurons that respond to noxious cold (Story et al., 2003) and in recombinant expression systems noxious cold has been shown to directly activate this channel (Sawada et al., 2007). Furthermore, the same set of sensory neurons in DRG and neurons in the trigeminal ganglia produce nociceptive responses to irritant agents such allyl isothiocyanate (AITC) from mustard oil, allicin from garlic and cinnamaldehyde from cinnamon that have all been shown to selectively activate TRPA1 (Bandell et al., 2004). TRPA1 deficient mice do not show pain behaviour associated with administration of AITC or noxious cold, clearly demonstrating that these behaviours are mediated by TRPA1 (Kwan et al., 2006).
Application Notes:
hTRPA1 channels have been selectively expressed in a HEK293 cell line and characterized in terms of their biophysical and pharmacological properties using whole-cell patch clamp techniques and Ca2+ fluorescence.
hTRPA1 currents were activated by AITC and produced an outwardly rectifying I/V relationship. AITC also dose-dependently increased the intracellular Ca2+ concentration (EC50 = 12 µM) and the effect of 37 µM AITC was blocked by hTRPA1 antagonists ruthenium red (IC50 = 222 µM) and camphor (IC50 approximately 10 mM). AITC-activated currents showed the expected dependence on extracellular Ca2+ typical of hTRPA1 channels.
Cinnamaldehyde (200 µM) also activated significant outward current at +80 mV (>2 nA) that was blocked by the hTRPA1 antagonist camphor (3 mM) using IonWorksTM automated electrophysiology. Cinnamaldehyde dose-dependently increased the intracellular Ca2+ concentration (EC50 = 16 µM) and the effect of 25 µM cinnamaldehyde was blocked by hTRPA1 antagonists ruthenium red (IC50 = 73 µM) and camphor (IC50 approximately 10 mM).
hTRPA1 current amplitudes, monitored using IonWorksTM HT, were consistently around 2-2.5 nA and percent expression levels above 80% (hTRPA1 currents >500 pA at +80 mV) for at least 29 cell passages.
These data demonstrate the selective and stable functional expression of TRPA1 channels in a HEK293 cell line.
hTRPA1 currents were activated by AITC and produced an outwardly rectifying I/V relationship. AITC also dose-dependently increased the intracellular Ca2+ concentration (EC50 = 12 µM) and the effect of 37 µM AITC was blocked by hTRPA1 antagonists ruthenium red (IC50 = 222 µM) and camphor (IC50 approximately 10 mM). AITC-activated currents showed the expected dependence on extracellular Ca2+ typical of hTRPA1 channels.
Cinnamaldehyde (200 µM) also activated significant outward current at +80 mV (>2 nA) that was blocked by the hTRPA1 antagonist camphor (3 mM) using IonWorksTM automated electrophysiology. Cinnamaldehyde dose-dependently increased the intracellular Ca2+ concentration (EC50 = 16 µM) and the effect of 25 µM cinnamaldehyde was blocked by hTRPA1 antagonists ruthenium red (IC50 = 73 µM) and camphor (IC50 approximately 10 mM).
hTRPA1 current amplitudes, monitored using IonWorksTM HT, were consistently around 2-2.5 nA and percent expression levels above 80% (hTRPA1 currents >500 pA at +80 mV) for at least 29 cell passages.
These data demonstrate the selective and stable functional expression of TRPA1 channels in a HEK293 cell line.
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:
hTRPA1
Presentation:
2 x 1 ml aliquots
Target Sub-Family:
TRP
Packaging:
2 x 1 ml aliquots
Protein or Enzyme Type:
Ion Channels
Product Name:
PrecisION hTRPA1-HEK Recombinant Cell Line