User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/15: Difference between revisions

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*** Well 6: 0.6 g/L Lysozyme
*** Well 6: 0.6 g/L Lysozyme
** Run as previously described. Refer to [http://openwetware.org/wiki/User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/09/25 Dr. Harting's protocol].
** Run as previously described. Refer to [http://openwetware.org/wiki/User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/09/25 Dr. Harting's protocol].
===Determination of molecular weight===
*By comparison with Precision Plus Protein Standards, the molecular weight of all samples was close to 15 KD, slightly lower, as expected for lysozyme compounds with MW=14,300 KD.
* There was a very small quantity of unknown A and colloid in the gel. The very faint mark observed in well 3 could even be considered to be bleed-over from the 30:1 colloid in well 4. Therefore, electrophoresis was inclonclusive with respect to the molecular weight of unknown A.
[[Image:Electrophoresis gel 151014.jpg]]
[[Image:Precision Plus Protein.jpeg|400x300px|]]
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Latest revision as of 00:26, 27 September 2017

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Tasks for October 15

  • To run SDS-PAGE
  • To perform data analysis of dialysed solutions from Oct. 14

SDS-PAGE

    • Warm your samples from Oct. 14 to about 40 °C using heating block
    • Transfer solutions to a premade gel
      • Well 1: Precision Plus Protein All Blue Standards
      • Well 3: 0.12 g/L lysozyme Ink A
      • Well 4: 30:1 Lysozyme colloid
      • Well 5: 0.12 g/L Lysozyme
      • Well 6: 0.6 g/L Lysozyme
    • Run as previously described. Refer to Dr. Harting's protocol.

Determination of molecular weight

  • By comparison with Precision Plus Protein Standards, the molecular weight of all samples was close to 15 KD, slightly lower, as expected for lysozyme compounds with MW=14,300 KD.
  • There was a very small quantity of unknown A and colloid in the gel. The very faint mark observed in well 3 could even be considered to be bleed-over from the 30:1 colloid in well 4. Therefore, electrophoresis was inclonclusive with respect to the molecular weight of unknown A.