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Lab Meeting August 8, 2013. By: Charis Fernandez. Techniques Learned. Gel Electrophoresis Cell culture Nanoparticle preparation Lyophilization Cytotoxicity. Native PAGE Bis-Tris Gel with Tricine Buffer. 24 hour run. 1 2 3 4 5 6 7. 1: Empty rHDL
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Lab Meeting August 8, 2013 By: Charis Fernandez
Techniques Learned • Gel Electrophoresis • Cell culture • Nanoparticle preparation • Lyophilization • Cytotoxicity
Native PAGE Bis-Tris Gel with Tricine Buffer 24 hour run 1 2 3 4 5 6 7 1: Empty rHDL 2: Myristoylvalrubicin 3: Apo A1 valrubicin 4: Thyroglobulin 5: Plasma serum 6: Bovine serum albumin 7: Human HDL Thyroglobulin MW: 660,000 Da BSA MW: 66,500 Da Human HDL MW: 70,000-360,000 Da Estimated rHDL MW: ~170,000 Da
Native PAGE Bis-TrisGel with Tricine Buffer 8 hour run 1 2 3 4 5 6 7 1: Empty rHDL 2: Myristoylvalrubicin 3: Apo A1 valrubicin 4: Thyroglobulin 5: Plasma Serum 6: Bovine Serum Albumin 7: Human HDL Thyroglobulin MW: 660,000 Da BSA MW: 66,500 Da Human HDL MW: 70,000-360,000 Da Estimated rHDL MW: ~170,000 Da
MTT Cytotoxicity Myr-5A-Cisx and Empty rHDL on SJ-N-KP cell line
Size and NMR to determine percent drug incorporated Empty Nanoparticle Samples • 0.5 mg Myr-5A peptide • 1.0 mg Myr-5A peptide • 2.0 mg Myr-5A peptide Encapsulated 50 µl Cisplatin Derivative Samples • 0.5 mg Myr-5A peptide • 1.0 mg Myr-5A peptide • 2.0 mg Myr-5A peptide
Future Studies • Western blot of Neuroblastomacell lines • Cytotoxicity of Cisplatin derivative on different Neuroblastoma cell lines • Effect of encapsulated Cisplatin derivative on normal healthy cells