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The clone N1E was subcloned by isolation of single cells on glass shards.

The adrenergic clone N1E-115 exhibits high levels of activity of the enzymes tyrosine hydroxylase and acetylcholinesterase which are necessary for neurotransmitter synthesis but are almost devoid of choline acetyltransferase.

These cells contain 14 receptors for neurotransmitters.

1. Remove medium, rinse with Modified Puck's Saline D1 solution:
   1. Glucose                       5.5 mM
   2. KCl                             5.4 mM
   3. Sucrose                     58.4 mM
   4. Na₂HPO₄-7H₂O         0.17 mM
   5. NaCl                          138 mM
   6. KH₂PO₄                     0.22 mM
   Adjusted to a pH of 7.4±0.05 with 0.1 N NaOH or 0.N HCl Adjusted to an osmomolality of 340±5 mosM with either sucrose or H₂0 to decrease or increase the osmomolality.
2. Allow flasks to remain at room temperature until cells detach.
3. To remove saline, add fresh culture medium, transfer cell suspension to centrifuge tube and spin at approximately 125 x for 5 to 10 minutes.
4. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
5. Place culture vessels in incubators at 37°C

Note: If DMEM formulation contain 1.5 g/L sodium bicarbonate.

. Methods in neurotransmitter receptor analysis. New York: Raven Press; 1990.

Richelson E. The use of cultured cells in the study of mood-normalizing drugs. Pharmacol. Toxicol. 66 suppl.3: 69-75, 1990. PubMed: 2179933

Amano T, et al. Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase- acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc. Natl. Acad. Sci. USA 69: 258-263, 1972. PubMed: 4400294

Richelson E. Regulation of tyrosine hydroxylase activity in mouse neuroblastoma clone N1E-115. J. Neurochem. 21: 1139-1145, 1973. PubMed: 4148612