User:James C. Schwabacher/Notebook/CHEM-571/2014/01/29: Difference between revisions

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## .0064 g of BSA in 10mL of water --> 9.69 μM
## .0064 g of BSA in 10mL of water --> 9.69 μM
#L-Cysteine
#L-Cysteine
## somethihe
## .0259 g in 25 mL water --> 8.55 mM
## something else --> 10μM
## 11.7 uL in 10 mL water --> 10μM
#Ammonium Hydroxide
#Ammonium Hydroxide
## 6.76 μL of 28-30% Ammonium Hydroxide in 10 mL
## 6.76 μL of 28-30% Ammonium Hydroxide in 10 mL

Revision as of 12:24, 29 January 2014

Biomaterials Design Lab <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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objective

The objective for today is to begin the synthesis of BSA:L-Cysteine Nanocrystals

Preparation of stock solutions

  1. BSA
    1. .0064 g of BSA in 10mL of water --> 9.69 μM
  2. L-Cysteine
    1. .0259 g in 25 mL water --> 8.55 mM
    2. 11.7 uL in 10 mL water --> 10μM
  3. Ammonium Hydroxide
    1. 6.76 μL of 28-30% Ammonium Hydroxide in 10 mL
  4. Sodium Sulfide

Protocol BSA:L-cysteine Nanocrystals

  1. In test tube mix 180 uL of 5mM Zinc Nitrate and 20 uL of 5mM Manganese Acetate
  2. Add 200 uL of 40 mM Ammonium Acetate
  3. Add 200 uL of Protein:ligand in various ratios from 0% to 100% protein
  4. Adjust pH to 8.2 using roughly 250 uL 1 mL of 10mM Ammonium Hydroxide, and bring volume of water to 2.3 mL
  5. Incubate at room temp for 1 hour
  6. Add 200 uL of 5mM Sodium Sulfide drop wise while vortexing
  7. Incubate for five days at 37 degrees Celsius

Table of BSA:L-Cysteine ratios

mol of protein 2.00E-09 1.80E-09 1.60E-09 1.40E-09 1.20E-09 1.00E-09 8.00E-10 6.00E-10 4.00E-10 2.00E-10 0
percentage of protein 100 90 80 70 60 50 40 30 20 10 0
percentage of ligand 0 10 20 30 40 50 60 70 80 90 100
mol of ligand 0 2.00E-10 4.00E-10 6.00E-10 8.00E-10 1.00E-09 1.20E-09 1.40E-09 1.60E-09 1.80E-09 2.00E-09

Notes

This area is for any observations or conclusions that you would like to note.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.




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