User:Khyra A. Neal/Notebook/Chem 571/2014/10/15: Difference between revisions

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'''NOTE''' The final concentration of NH<sub>4</sub>SCN was determined by titration standardization on [http://openwetware.org/wiki/User:Khyra_A._Neal/Notebook/Chem_571/2014/10/08 October 8, 2014]
'''NOTE''' The final concentration of NH<sub>4</sub>SCN was determined by titration standardization on [http://openwetware.org/wiki/User:Khyra_A._Neal/Notebook/Chem_571/2014/10/08 October 8, 2014]
'''Titrations were performed as follows'''
* 9.68 mM NH<sub>4</sub> is being titrated against 3 mL 1M HNO<sub>3</sub>, 10 mL deionized H<sub>2</sub>O , 200 μLFerric alum indicator, and 0.5 mL of dialyzed sample
** Lysozyme in 2 mM KI
*** 15.1 mL NH<sub>4</sub>SCN used
** Lysozyme in 5 mM KI
*** 14.2 mL NH<sub>4</sub>SCN used
** Lysozyme in 10 mM KI
*** 14.0 mL NH<sub>4</sub>SCN used
** Lysozyme in 25 mM KI
*** 13.7 mL NH<sub>4</sub>SCN used
** Lysozyme in 50 mM KI
*** 12.5 mL NH<sub>4</sub>SCN used




Revision as of 09:10, 21 October 2014

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October 15, 2014

Tasklist

Electrophoresis

  • Warm solutions for 5 minutes at 40°C that were prepared on October 14, 2014
  • Load samples to premade gel
    • Well 1: Precision Plus Protein All Blue Standards
    • Well 3: 0.12 g/L lysozyme Unk A
    • Well 4: 30:1 Lysozyme colloid
    • Well 5: 0.12 g/L Lysozyme
    • Well 6: 0.6 g/L Lysozyme
    • Wells 8-12 contain solutions prepared by Dr. Fox
  • Run for 40 minutes at 200V
  • Develop/Stain your gel
    • Place gel in Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes
    • Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour
    • Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes
      • Repeat this step with fresh destain solution 2 more times

The original protocol for electrophoresis was written by Dr. Hartings


Determination of Dialyzed KI Samples extracted on October 14, 2014

Determination of KI Concentration by Titration

  • Standardization:
    • AgNO3 + NH4SCN → AgSCN + NH4NO3
    • KI + AgNO3(excess) → AgI (s) + AgNO3 +KNO3
      • Initial AgNO3: 0.075 M AgNO3 x 0.002 L AgNO3 = 0.00015 mol AgNO3
      • AgNO3 (neutralized): 0.00968 M NH4SCN x 0.0104 L NH4SCN x (1 mol AgNO3 / 1 mol NH4SCN ) = 0.000100672 mol AgNO3
      • Moles of KI: 0.00015 mol AgNO3 initial - 0.000100672 mol AgNO3 neutralized = 0.0000493 mol KI
      • [KI]= 0.0000493 mol KI / 0.001 L KI = 49.3 mM KI

NOTE The final concentration of NH4SCN was determined by titration standardization on October 8, 2014


Titrations were performed as follows

  • 9.68 mM NH4 is being titrated against 3 mL 1M HNO3, 10 mL deionized H2O , 200 μLFerric alum indicator, and 0.5 mL of dialyzed sample
    • Lysozyme in 2 mM KI
      • 15.1 mL NH4SCN used
    • Lysozyme in 5 mM KI
      • 14.2 mL NH4SCN used
    • Lysozyme in 10 mM KI
      • 14.0 mL NH4SCN used
    • Lysozyme in 25 mM KI
      • 13.7 mL NH4SCN used
    • Lysozyme in 50 mM KI
      • 12.5 mL NH4SCN used







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