User:Trisha I. Ibeh/Notebook/Trisha Notebook/2013/10/23

From OpenWetWare

< User:Trisha I. Ibeh | Notebook | Trisha Notebook | 2013 | 10(Difference between revisions)
Jump to: navigation, search
(Procedure)
Current revision (06:55, 3 November 2013) (view source)
(Notes)
 
Line 76: Line 76:
*Fibers not formed
*Fibers not formed
-
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
+
 
 +
<u>Stock Solutions</u>
 +
# Gold
 +
## 49.3 mg HAuCl<sub>4</sub>·3H<sub>2</sub>O in 50mL water ---> 2.50mM
 +
# HRP
 +
## 6.6 mg HRP in 100 mL water ---> 1.5μM
 +
# Myoglobin
 +
## 28.7mg myoglobin in 100mL water ---> 16.9μM<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
|}
|}
__NOTOC__
__NOTOC__

Current revision

Biomaterials Design Lab Main project page
Previous entry      Next entry

Objective

  • Measure pepsin activity of pepsin in solution and pepsin-AuNPs in solution
  • Run UV-Vis on HRP and Pepsin linked Au-NP's synthesized yesterday
  • Synthesize HRP- AuNP at higher ratios.
  • Synthesize Myoglobin- AuNPs with the ratio used in yesterday's lab

Procedure

  1. Protein-AuNP analysis
    1. Take a picture and note physical description of each ratio
    2. Place 1mL of each ratio in an 1.5mL eppendorf tube.
    3. Centrifuge for 30 minutes (Be considerate of other uses here. Act quickly, and get as many tubes in the centrifuge as you can at once.)
    4. Take a UV-Vis spectrum of each sample (Be considerate of other users here)
    5. After taking the spectrum, save the sample in a fresh eppy tube.
  2. Synthesis of HRP-AuNPs and Hemoglobin-AuNPs
    1. Into test tube that will contain a total of 5mL add
    2. Au solution such that the final concentration is 0.25mM
    3. For Myoglobin make the following solutions to reflect an Au:Protein ratios
      1. 30
      2. 50
      3. 70
      4. 90
      5. 110
      6. 130
      7. 150
      8. 170
      9. 190
      10. 210
    4. For HRP make the following solutions to reflect an Au:Protein ratios
      1. 230
      2. 250
      3. 270
      4. 290
      5. 310
      6. 330
      7. 350
      8. 370
      9. 390
      10. 410
    5. Water up to 5mL
    6. Place solutions in oven for 4 hours at 80 degrees Celsius


Calculations used to make solutions

Image:Screen_Shot_2013-11-03_at_5.38.05_AM.png

Data

Image: Screen_Shot_2013-11-03_at_5.22.18_AM.png

Image: Screen_Shot_2013-11-03_at_5.23.38_AM.png

Image: Screen_Shot_2013-11-03_at_5.24.59_AM.png

Image:Screen_Shot_2013-11-03_at_5.26.21_AM.png

Notes

Observations

HRP

  • Purple tint of varying degrees:Lower ratios were magenta and higher ratios were blueish purple)
  • No fibers formed

Pepsin

  • Fibers not formed


Stock Solutions

  1. Gold
    1. 49.3 mg HAuCl4·3H2O in 50mL water ---> 2.50mM
  2. HRP
    1. 6.6 mg HRP in 100 mL water ---> 1.5μM
  3. Myoglobin
    1. 28.7mg myoglobin in 100mL water ---> 16.9μM


Personal tools