Bamboo splints, 6" |
Ted Pella Inc |
116 |
|
Black dissecting dish |
|
|
Glass petri dish filled to rim with SYLG170 or SYLG184 (colored black with finely ground charcoal powder). Leave at room temperature for 24-48 h to polymerize. |
Blade holder |
Fine Science Tools |
10053 |
|
Bovine serum albumin |
Sigma-Aldrich |
A7906 |
|
Double-sided tape |
3M |
665 |
|
Drosophila& food media, nutrient-rich |
|
|
7.5% sucrose, 15% glucose, 2.5% agar, 20% brewers yeast, 5% peptone, 0.125% MgSO4.7H2O, 0.125% CaCl2.2H20 |
Drosophila& food media, standard |
|
|
Bloomington Drosophila Stock center cornmeal recipe. (https://bdsc.indiana.edu/information/recipes/bloomfood.html) |
Fixative solution |
|
|
4% formadehyde in PBS, pH 7.4. |
Forceps |
Fine Science Tools |
91150-20 |
Forceps should be sharpened frequently. |
Formaldehyde |
Thermo Scientific |
28908 |
|
Glass 9-well dishes |
Corning |
7220-85 |
Also known as 9-well dishes |
Glass coverslips (22 x 22 mm) |
Fisher Scientific |
12-542-B |
|
Glass microscope slides (25 x 75 x 1 mm) |
Fisher Scientific |
12-550-413 |
|
Glass petri dish |
Corning |
3160-100BO |
|
Glycerol |
Sigma-Aldrich |
G5516 |
|
Microcentrigure tubes |
Axygen |
MCT-175-C |
|
Microdissection scissors |
Fine Science Tools |
15000-03 |
|
Microwell trays (72 x 10 µL wells) |
Nunc |
438733 |
|
Mounting media |
|
|
0.5% N-propylgallate and 80% glycerol in PBS |
N-propylgallate |
Sigma-Aldrich |
P3130 |
|
PBS (phosphate buffered saline pH 7.4) |
Sigma-Aldrich |
P5368 |
Prepare according to manufacturer's instructions |
PBT |
|
|
0.15% TritonX and 0.5% bovine serum albumin in PBS, pH 7.4 |
Pin holder |
Fine Science Tools |
26016-12 |
|
Primary antibody: goat anti-GAPDH |
Imgenex |
IMG-3073 |
For Western blotting. Used at 1:3000 |
Primary antibody: rabbit anti-cleaved Dcp-1 |
Cell signaling |
9578S |
For immunofluorescence. Used at 1:100 |
Primary antibody: rat anti-DEcad |
Developmental Studies Hybridoma Bank |
DCAD2 |
For immunofluorescence. Used at 1:20 |
Primary antibody: rat anti-DEcad |
DOI: 10.1006/dbio.1994.1287 |
DCAD1 |
Gift from Tadashi Uemura. Used at 1:100. |
Scalpel blades |
Fine Science Tools |
10050 |
Break off small piece of scapel blade and secure in blade holder. |
Secondary antibody: 488-conjugated donkey anti-rat IgG (H+L) |
Jackson ImmunoResearch |
712-545-153 |
For immunofluorescence. Used at 1:200 |
Secondary antibody: cy3-conjugated goat anti-rabbit IgG (H+L) |
Jackson ImmunoResearch |
111-165-144 |
For immunofluorescence. Used at 1:100 |
Secondary antibody: HRP-conjugated goat anti-rat IgG (H+L) |
Cell Signaling Technology |
7077 |
For Western blotting. Used at 1:3000 |
Secondary antibody: HRP-conjugated rabbit anti-goat IgG (H+L) |
Jackson ImmunoResearch |
305-035-003 |
For Western blotting. Used at 1:3000 |
Stereo dissecting microscope (M60 or M80) |
Leica Microsystems |
|
or similar microscope |
Sylgard (black) |
Dow Corning |
SYLG170 |
|
Sylgard (transparent) |
Dow Corning |
SYLG184 |
Color black with finely ground charcol powder |
Tissue: Kimwipes |
KIMTECH |
34120 |
|
TritonX |
Sigma-Aldrich |
T8787 |
|
Tungsten needle, fine |
Fine Science Tools |
10130-10 |
Insert into pin holder |
Tungsten needle, sturdy |
Fine Science Tools |
10130-20 |
Insert into pin holder |
Yeast paste |
(local supermarket) |
|
Approximately 2 tablespoons Fleischmann's ActiveDry Yeast (or similar) dissolved in ~20 mL distilled H2O |