The Rockefeller University
 SDS Agarose Gel Electrophoresis of Topo II
 Isomerase Poisons
 Student: Dr. Robert D. Craig...
 The Rockefeller University is a private
university which focuses primarily on basic
research in the biomedical fields an...
Rockefeller university
Who works here???
 Twenty-three Nobel Prize winners have been
associated with the university.
 The university has been t...
What happens when you Combine
soap with proteins?
 Both are zwitterions!
 Will form a clump! Or a miscelle
 Can use an ...
The surfactant surrounds DNA
fragments
sodium dodecylbenzene sulfonate
Sodium dodecylbenzene sulfonate -
Identification, toxicity, use,
water pollution potential...
sodium dodecy sulfonate
Sodium Dodecyl Sulfate
 (SDS) SDS is the most common dissociating
agent used to denature native proteins to
individual po...
Can surround an amino acid or “piece of
DNA”
SDS Gel electrophoresis
The uv transilluninator
Dr. Alfrey—SDS gel electrophoresis of mouse
leukemia cells treated with topo II isomerase
poisons
 It is similar to Paper...
This man, Dr. Alfrey, was really nice
and really famous!
 National Academy of Science
He would put me to sleep here
and there!
But, he took time out tell me some of
what he knew
This is what the famous guy told me!
 Topo II isomerase poisons are what your Mom
took!
 They are all we have right now!...
Cancer medications!
 Bleomycin
 Adriamycin
 Tamoxiphen Estrogen Hormone therapy!
He said “I study SDS Gel
electrophoresis”
You can go “in the Lab”
Check out what I do!
He said, “listen to everything I say”, if
you want to know about cancer!
I said, “I’ll listen!”
These Columns have electric fields to
migrate portions of DNA
SDS Gel electrophoresis: an Assay
SDS Gel electrophoresis: a series of
steps!
technique of
electrophoresis
 The distribution of charged species in a sample
can be shown experimentally by observing th...
The bowling lane!
technique of electrophoresis
 The solid structure of the matrix retards the
diffusion of the solute molecules, which will...
The matrix: Poly Acrylimide
(nail polish)
A hard gel! You can control the movement of
DNA, and amino acids
technique of electrophoresis
 At pH 6.00 alanine and isoleucine exist on
average as neutral zwitterionic molecules,
and a...
isoleucine
aspartic acid
arginine
Alanine
technique of electrophoresis
 The solute molecules of arginine therefore
carry an excess positive charge, and they
move t...
technique of electrophoresis
technique of
electrophoresis
The isoelectric points
The gel is “charged”
The amino acids are“charged”
Can separate proteins
Can also manipulate DNA
Intercalating Agent
 After we have removed our gel from the running
tank, we need to use ultraviolet light to visualize
t...
Intercalating Agent
 EtBr is an Intercalating Agent, meaning it
wedges itself into the grooves of DNA and stays
there. Mo...
And, detection with UV light
So, I can help those with cancer!
And, help kids play another day!
And, we can have some fun too!
Eating right!
Getting some excercise
No smoking! No drinking!
Or, meet the “five”
Sds presentation rockefeller[1][1]
Sds presentation rockefeller[1][1]
Upcoming SlideShare
Loading in …5
×

Sds presentation rockefeller[1][1]

800 views

Published on

Early Research of Topo II isomerase poisons
carried out by Dr. Alfrey at Rockefeller University

Published in: Education, Technology
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
800
On SlideShare
0
From Embeds
0
Number of Embeds
7
Actions
Shares
0
Downloads
18
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

Sds presentation rockefeller[1][1]

  1. 1. The Rockefeller University  SDS Agarose Gel Electrophoresis of Topo II  Isomerase Poisons  Student: Dr. Robert D. Craig, Ph.D  Mentor: Dr. Vincent Alfrey, M.D, Ph.D
  2. 2.  The Rockefeller University is a private university which focuses primarily on basic research in the biomedical fields and offers postgraduate and postdoctoral education. It is located between 63rd and 68th Streets along York Avenue, on the Upper East Side of Manhattan in New York City, New York. Its current president is Sir Paul Nurse.
  3. 3. Rockefeller university
  4. 4. Who works here???  Twenty-three Nobel Prize winners have been associated with the university.  The university has been the site of many important scientific breakthroughs. Rockefeller scientists, for example, established that DNA is the chemical basis of heredity, discovered blood groups, showed that viruses can cause cancer, founded the modern field of cell biology, worked out the structure of antibodies.
  5. 5. What happens when you Combine soap with proteins?  Both are zwitterions!  Will form a clump! Or a miscelle  Can use an electric field to guide down a silicon column or gel
  6. 6. The surfactant surrounds DNA fragments
  7. 7. sodium dodecylbenzene sulfonate Sodium dodecylbenzene sulfonate - Identification, toxicity, use, water pollution potential, ecological toxicity and regulatory information
  8. 8. sodium dodecy sulfonate
  9. 9. Sodium Dodecyl Sulfate  (SDS) SDS is the most common dissociating agent used to denature native proteins to individual polypeptides. When a protein mixture is heated to 100 °C in presence of SDS, the detergent wraps around the polypeptide backbone. It binds to polypeptides in a constant weight ratio of 1.4 g/g of polypeptide
  10. 10. Can surround an amino acid or “piece of DNA”
  11. 11. SDS Gel electrophoresis
  12. 12. The uv transilluninator
  13. 13. Dr. Alfrey—SDS gel electrophoresis of mouse leukemia cells treated with topo II isomerase poisons  It is similar to Paper chromatography!
  14. 14. This man, Dr. Alfrey, was really nice and really famous!  National Academy of Science
  15. 15. He would put me to sleep here and there!
  16. 16. But, he took time out tell me some of what he knew
  17. 17. This is what the famous guy told me!  Topo II isomerase poisons are what your Mom took!  They are all we have right now!  They stop all fast growing cells from replicating  That is why your Mom’s hair fell out  That is why your stomach gets upset  These medications stop fast reproducing tissue.
  18. 18. Cancer medications!  Bleomycin  Adriamycin  Tamoxiphen Estrogen Hormone therapy!
  19. 19. He said “I study SDS Gel electrophoresis”
  20. 20. You can go “in the Lab” Check out what I do!
  21. 21. He said, “listen to everything I say”, if you want to know about cancer!
  22. 22. I said, “I’ll listen!”
  23. 23. These Columns have electric fields to migrate portions of DNA
  24. 24. SDS Gel electrophoresis: an Assay
  25. 25. SDS Gel electrophoresis: a series of steps!
  26. 26. technique of electrophoresis  The distribution of charged species in a sample can be shown experimentally by observing the movement of solute molecules in an electric field, using the technique of electrophoresis. For such experiments an ionic buffer solution is incorporated in a solid matrix layer, composed of paper or a crosslinked gelatin-like substance. A small amount of the amino acid, peptide or protein sample is placed near the center of the matrix strip and an electric potential is applied at the ends of the strip,
  27. 27. The bowling lane!
  28. 28. technique of electrophoresis  The solid structure of the matrix retards the diffusion of the solute molecules, which will remain where they are inserted, unless acted upon by the electrostatic potential. In the example shown here, four different amino acids are examined simultaneously in a pH 6.00 buffered medium.
  29. 29. The matrix: Poly Acrylimide (nail polish)
  30. 30. A hard gel! You can control the movement of DNA, and amino acids
  31. 31. technique of electrophoresis  At pH 6.00 alanine and isoleucine exist on average as neutral zwitterionic molecules, and are not influenced by the electric field. Arginine is a basic amino acid. Both base functions exist as "onium" conjugate acids in the pH 6.00 matrix.
  32. 32. isoleucine
  33. 33. aspartic acid
  34. 34. arginine
  35. 35. Alanine
  36. 36. technique of electrophoresis  The solute molecules of arginine therefore carry an excess positive charge, and they move toward the cathode. The two carboxyl functions in aspartic acid are both ionized at pH 6.00, and the negatively charged solute molecules move toward the anode in the electric field.
  37. 37. technique of electrophoresis
  38. 38. technique of electrophoresis
  39. 39. The isoelectric points
  40. 40. The gel is “charged”
  41. 41. The amino acids are“charged”
  42. 42. Can separate proteins
  43. 43. Can also manipulate DNA
  44. 44. Intercalating Agent  After we have removed our gel from the running tank, we need to use ultraviolet light to visualize the DNA and verify that our Restriction Digest was successful. Does DNA fluoresce when exposed to UV light? Not on its own. Remember when we added dyes to our Blue-Juice to help us visualize the progress of the gel as it was running? We do something similar in this situation. Rather than "seeing" the DNA, we will be "seeing" a different molecule, called Ethidium Bromide (EtBr).
  45. 45. Intercalating Agent  EtBr is an Intercalating Agent, meaning it wedges itself into the grooves of DNA and stays there. More base pairs mean more gooves, which in turn means more EtBr can insert itself. This is an important concept. EtBr also has the property of fluorescing under UV light. So if we soak our gel in a solution of EtBr, it will intercalate into the DNA, then if we place our gel on or under a UV source, we can "see" the DNA by actually detecting the fluorescence of the EtBr. Wherever there is DNA, we will see a bright band at that point in the gel.
  46. 46. And, detection with UV light
  47. 47. So, I can help those with cancer!
  48. 48. And, help kids play another day!
  49. 49. And, we can have some fun too!
  50. 50. Eating right!
  51. 51. Getting some excercise
  52. 52. No smoking! No drinking!
  53. 53. Or, meet the “five”

×