1. Detection of Amnionless Protein in Kidney Sections Authors Natasha Tonge Dr. Roberta Rivi

2. Acknowledgements Dr. Roberta Rivi Dr. Sat Bhattacharya Dr. Katia Manova Dr. Betul Altay-Ozer Harlem Children Society Elizabeth Lacy Lab MSKCC

3. Wax in a Different Context Tissue sample is embedded in paraffin wax

4. Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome

5. Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Placed in Tissue bath

6. Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Placed in Tissue bath Mounted on a slide

7. Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Placed in Tissue bath Mounted on a slide Automatically Processed http://mskweb3.mskcc.org/MolecularCytology/mccf-final.htm

8. Immunohistochemistry - a process used to detect proteins in a tissue section. Antigen - a substance that provokes an immune response Antibody – the protein complex that binds to antigens Antibodies in a Different Context Enzyme - A protein that speeds up a chemical reaction. Substrate - The chemical that induces the chemical reaction

9. Immunohistochemistry - a process used to detect proteins in a tissue section. Antigen - a substance that provokes an immune response Antibody – the protein complex that binds to antigens Antibodies in a Different Context primary antibody secondary antibody substrate enzyme Antigen Enzyme - A protein that speeds up a chemical reaction. Substrate - The chemical that induces the chemical reaction

10. The Anatomy of the Kidney Proximal Tubule - close to the glomerulus. Ammnionless - protein located on the proximal tubule. Cortex - outer region of the kidney. Most likely to find the glomerulus and proximal tubule here. Medula - inner region of the kidney. Lacks ammnionless. http://upload.wikimedia.org/wikipedia/en/thumb/b/b0/Kidney_secti on.jpg/180px-Kidney_section.jpghttp://upload.wikimedia.org/wikipedia/en/thumb/b/b0/Kidney_secti on.jpg/180px-Kidney_section.jpg altered by Natasha Tonge

11. The Kidney in Focus (more or less)

13. The Past and Future of this Experiment Tissue samples are sectioned, placed on slides and stained The staining process, immunohistochemistry, makes use of antibodies to induce a color change In future experiments the same process will be used to pinpoint megalin and cubulin in abnormal mouse tissue. The results will then be accompanied by Western Blotting, Urine analysis and ELISA tests to characterize LRP Y2721- stop mice (mutants with shortened megalin sequences) To Detect Amnionless in Kidney Sections: That color change enabled us to see the location of the proximal tubules in the kidney sections.

14. Bibliography "Human Hair." Wikipedia. http://en.wikipedia.org/wiki/Human_ hair. 10 July 2006 http://en.wikipedia.org/wiki/Human_ Barnum, Susan R.. Biotechnology: An Introduction. 2. Belmont, CA: Thomson Brooks/Cole, 2005. Campbell A., Neil, Jane B. Reece, Lawrence G. Mitchell, and Martha R. Taylor.Biology: Concepts & Connections. 4th ed. San Fransisco: Benjamin Cummings, 2003.

15. owari