| 3-amino benzoic acid ethyl ester (Tricaine) powder |
Sigma-Aldrich |
A5040 |
Make 0.4% (15 mM) tricaine stock : Bring 0.4 g tricaine to 100 mL with egg water. Adjust to pH 7. |
| Dimethyl sulfoxide (DMSO) |
Sigma-Aldrich |
D8418 |
|
| Metronidazole |
Sigma-Aldrich |
M1547 |
|
| Triton X-100 |
Sigma-Aldrich |
T8532 |
|
| Formaldehyde |
Sigma-Aldrich |
252549 |
|
| Calcium sulfate dihydrate |
Sigma-Aldrich |
C3771 |
|
| N-Phenylthiourea (PTU) |
Sigma-Aldrich |
P7629 |
|
| Mounting media (Vectashield) |
Vector Laboratories |
H-1000 |
|
| 100 × 15 mm petri dish |
Denville Scientific Inc. |
M5300 |
|
| clear nail polish |
Fisher Scientific |
50949071 |
|
| fine forceps |
Fine Science Tools |
11251-20 |
Dumont #5 |
| glass slide |
Fisher Scientific |
12-544-1 |
|
| glass coverslip |
Fisher Scientific |
12-540-A and 12-542-A |
18 × 18 mm |
| zn-5 (mouse anti-Alcam) |
ZIRC |
zn-5 |
1:10 dilution |
| goat anti-Hnf4a |
Santa Cruz |
sc-6556 (C-19) |
1:50 dilution |
| rat anti-RFP |
Allele Biotechnology |
ACT-CM-MRRFP10 |
1:300 dilution |
| AlexaFluor 647 AffiniPure Donkey Anti-goat IgG (H+L) |
Jackson laboratories |
705-605-147 |
1:500 dilution |
| AlexaFluor 647 AffiniPure Goat Anti-mouse IgG (H+L) |
Invitrogen |
A21240 |
1:500 dilution |
| Cy3 donkey anti-rat IgG |
Jackson laboratories |
712-165-150 |
1:500 dilution |
| dissecting microscope |
Leica Microsystems |
S6F |
|
| epifluorescence microscope |
Leica Microsystems |
M205FA |
|
| confocal microscope |
Carl Zeiss |
LSM700 |
|
| egg water |
|
|
0.3 g/L of sea salts ‘Instant Ocean’ and 0.5 mM CaSO4 in distilled water. |
|
|
|
25.6 g Na2HPO4·7H2O, 80 g NaCl, 2 g KCl, 2 g KH2PO4. Bring to 1 L with distilled water. Adjust pH 7. |
| 10X PBS |
|
|
Autoclave for 20 min at 121°C. |
| 1X PBSDT |
|
|
0.2% Triton X-100 and 0.2% DMSO in 1 X PBS. |
| PEM buffer |
|
|
30.2 g PIPES, 761 mg EGTA, 1 mM MgSO4. Bring to 1 L with distilled water. Adjust pH 7. |
| Blocking solution |
|
|
Mix 1 ml of heat-inactivated horse serum with 9 ml of 1X PBSDT. |
