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Lab Meeting August 8, 2013

During the lab meeting held on August 8, 2013, Charis Fernandez presented critical techniques, including Gel Electrophoresis, cell culture, nanoparticle preparation, and lyophilization. Key findings included the cytotoxicity assays of various substances on the SJ-N-KP cell line, assessing the effects of Myristoylvalrubicin and empty rHDL nanoparticles. The presentation also highlighted the molecular weights of proteins involved, such as Thyroglobulin and BSA, and outlined future studies focusing on the impact of encapsulated Cisplatin derivatives on neuroblastoma cell lines and healthy cells.

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Lab Meeting August 8, 2013

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Presentation Transcript

  1. Lab Meeting August 8, 2013 By: Charis Fernandez

  2. Techniques Learned • Gel Electrophoresis • Cell culture • Nanoparticle preparation • Lyophilization • Cytotoxicity

  3. 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

  4. 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

  5. Myr-5A-Cisx Nanoparticle Size

  6. MTT Cytotoxicity Myr-5A-Cisx and Empty rHDL on SJ-N-KP cell line

  7. 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

  8. 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

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