User:Andrew K Farag/Notebook/Andrewfarag/2014/01/29

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Current revision (12:56, 29 January 2014) (view source)
 
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==objective==
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The objective for today is to begin the synthesis of BSA:L-Cysteine Nanocrystals
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==Objective==
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==Preparation of stock solutions==
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The objective for today was to make all stock solutions necessary for the BSA:L-cysteine Nanocrystals
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# BSA
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#L-Cysteine
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#Sodium Hydroxide
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#Sodium Sulfide
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==Protocol BSA:L-cysteine Nanocrystals==
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# In test tube mix 180 uL of 5mM Zinc Nitrate and 20 uL of 5mM Manganese Acetate
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# Add 200 uL of 40 mM Ammonium Acetate
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# Add 200 uL of Protein:ligand in various ratios from 0% to 100% protein
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# Adjust pH to 8.2 using roughly 250 uL of 10mM Ammonium Hydroxide, and bring volume of water to 2.3 mL 
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# Incubate at room temp for 1 hour
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# Add 200 uL of 5mM Sodium Sulfide drop wise while vortexing
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# Incubate for five days at 37 degrees Celsius
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==Table of BSA:L-Cysteine ratios==
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==Materials==
 
{| {{table}}
{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Name'''
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| 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
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| align="center" style="background:#f0f0f0;"|'''Formula'''
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| align="center" style="background:#f0f0f0;"|'''FW (g/mol)'''
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|-
|-
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| Zinc Nitrate, 6-Hydrate Crystal||Zn(NO3)2 * 6H2O||297.47
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| percentage of protein||100||90||80||70||60||50||40||30||20||10||0
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|-
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| Manganese (II) acetate tetrahydrate (good for 2 weeks)||Mn(CH3COO)2 * 4H2O||245.08
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| percentage of ligand||0||10||20||30||40||50||60||70||80||90||100
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|-
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| Ammonium Acetate||NH4(CH3COO)||77.08
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| 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
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|-
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| Sodium Sulfide nonahydrate(fresh stock) ||Na2S * 9 H2O||240.18
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|-
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| L-Cysteine (2 weeks)||HSCH2CH(NH2)CO2H||121.16
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|-
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| BSA (fresh stock)||||66kDa
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|-
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| Ammonium Hydroxide||NH4OH||14.8M (6.76uL in 10 mL)
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|}
|}
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==Preparation of Stocks==
 
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# 5mM Zinc Nitrate
 
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# 5.55mM Manganese Acetate
 
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# 41.65 mM Ammonium Acetate
 
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*0.0066g in 10mL BSA will be made tomorrow
 
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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
  2. L-Cysteine
  3. Sodium Hydroxide
  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 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 protein2.00E-091.80E-091.60E-091.40E-091.20E-091.00E-098.00E-106.00E-104.00E-102.00E-100
percentage of protein1009080706050403020100
percentage of ligand0102030405060708090100
mol of ligand02.00E-104.00E-106.00E-108.00E-101.00E-091.20E-091.40E-091.60E-091.80E-092.00E-09



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