This solution will be in dialysis with the cytosol and hence is similar to it in composition (high potassium and low sodium).
| Chemical |
mM |
MW/concentration |
for 100 ml |
| KCl |
107 |
74.55 |
0.7976 g |
| CaCl2 |
1 |
1 M |
100 ul |
| MgCl2 |
7 |
1 M |
700 ul |
| EGTA |
11 |
380.35 |
0.4183 |
| HEPES |
10 |
238.3 |
0.2383 g |
| Na2ATP |
5 |
569.16 |
0.2846 |
Adjust to pH to 7.2 using potassium hydroxide (KOH).
Always check the molecular weight on the bottles of the chemicals. It may vary, due to different water contents for example.
Reference
Chimerel et al., Bacterial metabolite indole modulates incretin secretion from intestinal enteroendocrine L cells, Cell Rep., 2014
Link to the article in PubMed
This solution is needed to keep cells in a physiological environment during patch-clamp experiments. It also contains glucose as a nutrient.
| Chemical |
mM |
MW/concentration |
for 1 L |
for 500 ml |
for 500 ml 10X |
| NaCl |
138 |
58.44 |
8.065 g |
4.0325 g |
40.325 g |
| KCl |
4.5 |
74.55 |
0.335 g |
0.1675 g |
1.675 g |
| NaHCO3 |
4.2 |
84.01 |
0.3528 g |
0.1764 g |
1.764 g |
| NaH2PO4 |
1.2 |
120 |
0.144 g |
0.072 g |
0.72 g |
| CaCl2 |
2.6 |
1 M |
2.6 ml |
1.3 ml |
– |
| MgCl2 |
2.6 |
1 M |
1.2 ml |
0.6 ml |
6 ml |
| HEPES |
10 |
238.3 |
2.382 g |
1.1915 g |
11.915 g |
Glucose: 1 mM (18 MG/100 ml)
pH 7.4 using NaOH
For a 500 ml 1X solution, I added 2.4 ml of NaOH at 1 M concentration.
Add CaCl2 and adjust the pH when the solution concentration is 1X.
Add glucose to the solution needed on the day, usually 50 ml.
Always check the molecular weight on the bottles of the chemicals. It may vary, due to different water contents for example.
Reference
Rogers et al., Electrical activity-triggered glucagon-like peptide-1
secretion from primary murine L-cells, J. Physiol., 2011
Link to the article in PubMed
Activités scientifiques de Cristian Riccio