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
Cette solution est utilisée après avoir coloré un gel de polyacrylamide au bleu de Coomassie. Il faut faire baigner le gel dans cette solution, chauffer 1 min au micro-ondes et laisser décolorer sur une plateforme en mouvement pendant quelques dizaines de minutes. Ensuite, jeter le liquide coloré dans l’évier et répéter jusqu’à obtention d’un gel suffisamment décoloré (jusqu’à ce que seules les bandes de protéines restent bleues).
Pour 2 litres:
– 1 litre d’EtOH (éthanol pur ou éthanol avec 5% de méthanol)
– 800 ml d’H20
– 200 ml d’acide éthanoïque (= acide acétique)
Si l’acide éthanoïque est à 80% -> verse 850 ml d’H20 et 250 ml d’acide éthanoïque à la place.
Activités scientifiques de Cristian Riccio