User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/23

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==Purpose==
==Purpose==
* Clean up and make room for samples of Lysozyme/Gold solutions
* Clean up and make room for samples of Lysozyme/Gold solutions
-
* Make gold/lysozyme solutions in with the following ratios:  
+
* Make stock solutions for Luminol assay experiment tomorrow: Luminol, Horseradish peroxidase, H2O2, sodium carbonate buffer.
-
      10-15-20-25-30-35-40
+
==Procedure==
==Procedure==
 +
* Test tubes and falcon tubes were washed for clean up.
 +
* A serious of stocks solutions were made for testing luminol experiment the next day.
 +
**Phenol: concentration of 18mM 4-Iodophenol was not made because about 18mM phenol stock solution pre-made from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/18|2012/09/18]]  (check this data entry for calculations made and actual concentration of phenol) is going to be used for luminol assay due to its long shelf life in room temperature.
 +
**Horseradish peroxidase
 +
    Horseradish peroxidase was made by weighting out 0.004g of horseradish peroxidase power.
 +
    The weighted out amount was mixed with 1mL of sterile water.
 +
    Molecular weight of horseradish peroxidase is 66kDa.
 +
    Calculation is depicted as follows:
 +
    0.004g X 66,000g/mol ÷ 0.001L = 9.2X10<sup>-6</sup>M = 9.2μM
 +
**Carbonate buffer
 +
    Carbonate buffer was made by mixing 100mL of 1M sodium bicarbonate with 100mL of 1M sodium bicarbonate together.
 +
    pH of carbonate buffer was diluted by adding 1M and 6M HCl into the solution
 +
    For calculations done to obtain the amount of grams weighted out the sodium carbonate and sodium bicarbonate, please consult [[User:Melissa Novy/Notebook/CHEM-571/2012/10/23|Melissa's Notebook]].
 +
    Molecular weight of sodium carbonate: 105.9784g/mol
 +
    0.0106g X 105.9784g/mol ÷ 0.1L = 1X10<sup>-3</sup>M = 1mM sodium carbonate
 +
 +
    Molecular weight of sodium bicarbonate: 84.007g/mol
 +
    0.0084g X 84.007g/mol ÷ 0.1L = 1X10<sup>-3</sup>M = 1mM sodium bicarbonate
 +
 +
   

Revision as of 12:25, 26 October 2012

Experimental Biological Chemistry I Main project page
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Purpose

  • Clean up and make room for samples of Lysozyme/Gold solutions
  • Make stock solutions for Luminol assay experiment tomorrow: Luminol, Horseradish peroxidase, H2O2, sodium carbonate buffer.

Procedure

  • Test tubes and falcon tubes were washed for clean up.
  • A serious of stocks solutions were made for testing luminol experiment the next day.
    • Phenol: concentration of 18mM 4-Iodophenol was not made because about 18mM phenol stock solution pre-made from 2012/09/18 (check this data entry for calculations made and actual concentration of phenol) is going to be used for luminol assay due to its long shelf life in room temperature.
    • Horseradish peroxidase
   Horseradish peroxidase was made by weighting out 0.004g of horseradish peroxidase power. 
   The weighted out amount was mixed with 1mL of sterile water.
   Molecular weight of horseradish peroxidase is 66kDa.
   Calculation is depicted as follows:
   0.004g X 66,000g/mol ÷ 0.001L = 9.2X10-6M = 9.2μM
    • Carbonate buffer
   Carbonate buffer was made by mixing 100mL of 1M sodium bicarbonate with 100mL of 1M sodium bicarbonate together.
   pH of carbonate buffer was diluted by adding 1M and 6M HCl into the solution
   For calculations done to obtain the amount of grams weighted out the sodium carbonate and sodium bicarbonate, please consult Melissa's Notebook.
   Molecular weight of sodium carbonate: 105.9784g/mol
   0.0106g X 105.9784g/mol ÷ 0.1L = 1X10-3M = 1mM sodium carbonate
   Molecular weight of sodium bicarbonate: 84.007g/mol
   0.0084g X 84.007g/mol ÷ 0.1L = 1X10-3M = 1mM sodium bicarbonate




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