Common experimental solutions
| Solution . | Composition (mM) . | Reference . |
|---|---|---|
| Relaxing | Kprop (170), magnesium acetate (2.5), MOPS (20), K2EGTA (5), and Na2ATP (2.5) | (Hessel et al., 2019) |
| Washing | Kprop (185), magnesium acetate (2.5), MOPS (20), and Na2ATP (2.5) | (Hessel et al., 2019) |
| Activating | Kprop (170), magnesium acetate (2.5), MOPS (10), Na2ATP (2.5), and different proportions of CaEGTA and K2EGTA to obtain target pCa | (Hessel et al., 2019) |
| Relaxing | KCl (100), imidazole (10), MgCl2 (1), EGTA (2), and Na2ATP (4.46) | (Jee and Lim 2016) |
| Activating | BES (40), CaCO3-EGTA (10), MgCl2 (6.29), Na2ATP (6.12), and Kprop (45.3); free Ca2+ set to pCa 4.0 | (Jee and Lim, 2016) |
| Relaxing | BES (40), EGTA (10), MgCl2 (6.56), Na2ATP (5.88), Kprop (46.35), and DTT (1) | (Tonino et al., 2017) |
| Activating | BES (40), CaCO3-EGTA (10), MgCl2 (6.29), Na2ATP (6.12), Kprop (45.3), and DTT (1) | (Tonino et al., 2017) |
| Relaxing | KCl (100), imidazole (20), MgATP (4), EGTA (2), and free Mg2+ (1) | (Fitzsimons et al., 2001) |
| Activating | KCl (79.2), imidazole (20), EGTA (7), MgATP (4), and free Mg2+ (1); free Ca2+ set to pCa 9.0-4.5 | (Fitzsimons et al., 2001) |
| Activating | Imidazole (20), EGTA (7), MgATP (4); free Mg2+ (1), and KCl; free Ca2+ set to pCa 4.5-9.0 | (Ochala et al., 2011) |
| Rigor | KCL (60), MgCl2 (5), MOPS (10), and EGTA (1) | (Suzuki and Ishiwata, 2011) |
| Relaxing | TES (100), MgCl2 (7.7), Na2ATP (5.44), EGTA (25), Na2CP (19.11), and GSH (10) | (Linari et al., 2007) |
| Washing | TES (100), MgCl2 (6.93), Na2ATP (5.45), EGTA (0.1), Na2CP (19.49), HDTA (24.9), and GSH (10) | (Linari et al., 2007) |
| Activation | TES (100), MgCl2 (6.76), Na2ATP (5.49), Na2CP (19.49), GSH (10), and CaEGTA (25) | (Linari et al., 2007) |
| Rigor | TES (100), MgCl2 (3.22), EGTA (53), and GSH 10 | (Linari et al., 2007) |
| Solution . | Composition (mM) . | Reference . |
|---|---|---|
| Relaxing | Kprop (170), magnesium acetate (2.5), MOPS (20), K2EGTA (5), and Na2ATP (2.5) | (Hessel et al., 2019) |
| Washing | Kprop (185), magnesium acetate (2.5), MOPS (20), and Na2ATP (2.5) | (Hessel et al., 2019) |
| Activating | Kprop (170), magnesium acetate (2.5), MOPS (10), Na2ATP (2.5), and different proportions of CaEGTA and K2EGTA to obtain target pCa | (Hessel et al., 2019) |
| Relaxing | KCl (100), imidazole (10), MgCl2 (1), EGTA (2), and Na2ATP (4.46) | (Jee and Lim 2016) |
| Activating | BES (40), CaCO3-EGTA (10), MgCl2 (6.29), Na2ATP (6.12), and Kprop (45.3); free Ca2+ set to pCa 4.0 | (Jee and Lim, 2016) |
| Relaxing | BES (40), EGTA (10), MgCl2 (6.56), Na2ATP (5.88), Kprop (46.35), and DTT (1) | (Tonino et al., 2017) |
| Activating | BES (40), CaCO3-EGTA (10), MgCl2 (6.29), Na2ATP (6.12), Kprop (45.3), and DTT (1) | (Tonino et al., 2017) |
| Relaxing | KCl (100), imidazole (20), MgATP (4), EGTA (2), and free Mg2+ (1) | (Fitzsimons et al., 2001) |
| Activating | KCl (79.2), imidazole (20), EGTA (7), MgATP (4), and free Mg2+ (1); free Ca2+ set to pCa 9.0-4.5 | (Fitzsimons et al., 2001) |
| Activating | Imidazole (20), EGTA (7), MgATP (4); free Mg2+ (1), and KCl; free Ca2+ set to pCa 4.5-9.0 | (Ochala et al., 2011) |
| Rigor | KCL (60), MgCl2 (5), MOPS (10), and EGTA (1) | (Suzuki and Ishiwata, 2011) |
| Relaxing | TES (100), MgCl2 (7.7), Na2ATP (5.44), EGTA (25), Na2CP (19.11), and GSH (10) | (Linari et al., 2007) |
| Washing | TES (100), MgCl2 (6.93), Na2ATP (5.45), EGTA (0.1), Na2CP (19.49), HDTA (24.9), and GSH (10) | (Linari et al., 2007) |
| Activation | TES (100), MgCl2 (6.76), Na2ATP (5.49), Na2CP (19.49), GSH (10), and CaEGTA (25) | (Linari et al., 2007) |
| Rigor | TES (100), MgCl2 (3.22), EGTA (53), and GSH 10 | (Linari et al., 2007) |
Recipes common in the field. Omitted is the addition of creatine phosphate (∼15 mM) and creatine kinase (400–500 U/ml), which is typically added to all but rigor solutions to replenished ATP; however, for experiments <2 h, we find >2.5 mM ATP lasts without problem. Finally, the addition of protease inhibitors to all solutions can limit protein degradation (see text). All solutions can be set to a pH 7–7.1 at the experimental temperature. BES, N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid; EGTA, ethylene glycol bis (β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid; CrP, creatine phosphate; CK, creatine kinase; GSH, glutathione; DTT, dithiothreitol; HDTA, 1,6-diaminohexane-N,N,N′,N′-tetraacetic acid; MOPS, 3-(N-morpholino)propanesulfonic acid; TES, N-[Tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid; Kprop, potassium propionate.