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Detection of Amnionless Protein in Kidney Sections. Authors Natasha Tonge Dr. Roberta Rivi. Acknowledgements. Dr. Roberta Rivi Dr. Sat Bhattacharya Dr. Katia Manova Dr. Betul Altay-Ozer Harlem Children Society Elizabeth Lacy Lab MSKCC. Wax in a Different Context. - PowerPoint PPT Presentation
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Detection of Amnionless Protein in
Kidney Sections
AuthorsNatasha TongeDr. Roberta Rivi
Acknowledgements
Dr. Roberta RiviDr. Sat Bhattacharya
Dr. Katia ManovaDr. Betul Altay-Ozer
Harlem Children SocietyElizabeth Lacy Lab
MSKCC
Wax in a Different Context
Tissue sample is embedded in paraffin wax
Wax in a Different Context
Tissue sample embedded in paraffin wax
Sliced with microtome
Wax in a Different Context
Tissue sample embedded in paraffin waxSliced with microtome
Placed in Tissue bath
Wax in a Different Context
Tissue sample embedded in paraffin waxSliced with microtomePlaced in Tissue bath
Mounted on a slide
Wax in a Different Context
Tissue sample embedded in paraffin waxSliced with microtomePlaced in Tissue bathMounted on a slide
Automatically Processed
http://mskweb3.mskcc.org/MolecularCytology/mccf-final.htm
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
Immunohistochemistry - a process used to detect proteins in a tissue section.Antigen - a substance that provokes an immune responseAntibody – 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
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_section.jpg/180px-Kidney_section.jpg altered by Natasha Tonge
The Kidney in Focus (more or less)
The Kidney in Focus (more or less)
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 topinpoint 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.
Bibliography
"Human Hair." Wikipedia. http://en.wikipedia.org/wiki/Human_hair. 10 July 2006
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.
owari
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