7. Solution of trypsin (0.25%) and ethylenediamine tetraacetic
acid (EDTA).
8. Immortal Mouse Fibroblast Cell Line (3T3; ATCC).
9. 16% Paraformaldehyde.
10. FACS Fixation Buffer: 10 mL of 16% PFA diluted with 30 mL
of PBS. Solution should be stored with light protection at 4
C.
11. Live/Dead Fixable Aqua Dead Cell Stain, 405 nm excitation.
This dye is reconstituted in DMSO and is stored at À20
C,
light protected, when not in use.
12. Dimethyl Sulfoxide (DMSO).
13. Cy5.5-CTP-amide: Prepared by peptide synthesis, conjugated
with Cy5.5 fluorophore, and purified using HPLC. Lyophilized powder is reconstituted in DMSO at 10 mM concentration. After reconstituting, store at -80
C.
14. Glass-bottomed dishes (MatTek).
2.1.2 Bio-distribution
In Vivo
1. 6- to 12-week-old albino, female mice (CD1, Charles River).
2. 28G 0.5 mL Insulin syringes.
3. 100 mg/mL Ketamine HCl (KetaVed).
4. 20 mg/mL Xylazine (AnaSed).
5. Ketamine/Xylazine Solution: Ketamine HCl is mixed with
Xylazine 1:1 to produce a stock solution containing 50 mg/
mL ketamine and 10 mg/mL Xylazine. This solution is made
fresh before each use.
6. 26G Needles.
7. 3 mL Syringe.
8. 10% Buffered Formalin Phosphate.
9. Paraplast X-TRA (SIGMA).
10. Ethanol.
11. Xylene.
12. 10Â Tris Buffered Saline (TBS).
13. 1Â TBS Solution: 10Â TBS is diluted 1:10 with deionized
water. This solution can be stored at room temperature.
14. Microscope Slides.
15. Cover Glass.
16. Dapi Fluoromount G (SouthernBiotech).
17. Sparkle Optical Lens Cleaner.
18. Tissue-Tek Processing/Embedding Cassettes (ThermoFisher).
Validating Cardiac Targeting Peptide
101
acid (EDTA).
8. Immortal Mouse Fibroblast Cell Line (3T3; ATCC).
9. 16% Paraformaldehyde.
10. FACS Fixation Buffer: 10 mL of 16% PFA diluted with 30 mL
of PBS. Solution should be stored with light protection at 4
C.
11. Live/Dead Fixable Aqua Dead Cell Stain, 405 nm excitation.
This dye is reconstituted in DMSO and is stored at À20
C,
light protected, when not in use.
12. Dimethyl Sulfoxide (DMSO).
13. Cy5.5-CTP-amide: Prepared by peptide synthesis, conjugated
with Cy5.5 fluorophore, and purified using HPLC. Lyophilized powder is reconstituted in DMSO at 10 mM concentration. After reconstituting, store at -80
C.
14. Glass-bottomed dishes (MatTek).
2.1.2 Bio-distribution
In Vivo
1. 6- to 12-week-old albino, female mice (CD1, Charles River).
2. 28G 0.5 mL Insulin syringes.
3. 100 mg/mL Ketamine HCl (KetaVed).
4. 20 mg/mL Xylazine (AnaSed).
5. Ketamine/Xylazine Solution: Ketamine HCl is mixed with
Xylazine 1:1 to produce a stock solution containing 50 mg/
mL ketamine and 10 mg/mL Xylazine. This solution is made
fresh before each use.
6. 26G Needles.
7. 3 mL Syringe.
8. 10% Buffered Formalin Phosphate.
9. Paraplast X-TRA (SIGMA).
10. Ethanol.
11. Xylene.
12. 10Â Tris Buffered Saline (TBS).
13. 1Â TBS Solution: 10Â TBS is diluted 1:10 with deionized
water. This solution can be stored at room temperature.
14. Microscope Slides.
15. Cover Glass.
16. Dapi Fluoromount G (SouthernBiotech).
17. Sparkle Optical Lens Cleaner.
18. Tissue-Tek Processing/Embedding Cassettes (ThermoFisher).
Validating Cardiac Targeting Peptide
101