Formation method of inside-out patch in patch clamp experiment
An inside-out, experimental method technology, applied in the field of inside-out patch formation of the patch clamp, to achieve the effect of improving experimental efficiency, simple operation, and increasing success rate
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[0031] Hereinafter, taking HEK-293 cells recombinantly expressing the large conductance calcium-activated potassium channel (BKCa) of human vascular smooth muscle as an example, the operation method and steps of the present invention will be described in detail. However, it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.
[0032] 1. Liquid preparation
[0033] Electrode solution: D-aspartic acid40mM, KOH40mM, KCl100mM, HEPES10mM, CaCl 2 1mM.
[0034] Low calcium bath: D-aspartic acid100mM, KOH100mM, KCl40mM, HEPES10mM, EGTA1mM, free Ca 2+ Concentration 10 -5 mM.
[0035] High calcium bath: D-aspartic acid100mM, KOH100mM, KCl40mM, HEPES10mM, EGTA1mM, free Ca 2+ Concentration 10 -4 mM.
[0036] Add 10mM Glucose to all liquids, adjust the osmotic pressure to 284mOSM, a...
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