User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/28: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
No edit summary
Line 29: Line 29:


===0.6 g/L lysozyme vs CaCl<sub>2</sub> using 3500 MWCO dialysis data===
===0.6 g/L lysozyme vs CaCl<sub>2</sub> using 3500 MWCO dialysis data===
<u>Bradford Analysis</u>
*Only running bradford analysis of protein-containing solutions, that is Lys 1, 2, 3, 4 and 5, since the protein cannot diffuse through 3500 MWCO.
* Remove 50 μL of solution from each chamber (7 in all) and run Bradford analysis
**Bradford reagent should be diluted 1:3 with 50mM Tris/50mM NaCl
**Recall, 50 μL solution + 200 μL diluted Bradford + 750 μL Tris/NaCl buffer
**PS cuvettes, measuring 400 - 800 nm
<u>Ca2+ ISE</u>
<u>Ca2+ ISE</u>
{| {{table}}
{| {{table}}

Revision as of 12:05, 29 October 2014

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Tasks for October 28

  • To continue analysis of dialysed 0.6 g/ L Lysozyme vs KI using 3500 MWCO
  • To check I- ISE values obtained on OCt. 21 and measure KI content of dialysed solutions
  • Set up dialysis of 0.6 g/L lysozyme vs CaCl2 using 3500 MWCO

I- ISE

KI concentration mV measurement (mV) (21 Oct.) mV measurement (mV) (28 Oct.)
2mM -198.3 -192.8
5 mM -217.3 -206.8
10 mM -237.0 -224.2
25 mM -247.4 -253.0
50mM -278.0 -276.2

0.6 g/L lysozyme vs CaCl2 using 3500 MWCO dialysis data

Bradford Analysis

  • Only running bradford analysis of protein-containing solutions, that is Lys 1, 2, 3, 4 and 5, since the protein cannot diffuse through 3500 MWCO.
  • Remove 50 μL of solution from each chamber (7 in all) and run Bradford analysis
    • Bradford reagent should be diluted 1:3 with 50mM Tris/50mM NaCl
    • Recall, 50 μL solution + 200 μL diluted Bradford + 750 μL Tris/NaCl buffer
    • PS cuvettes, measuring 400 - 800 nm

Ca2+ ISE

CaCl2 concentration mV measurement (mV)
5μM (1) 11.8
Lys (1) 4.6
50μM (2) 4.2
Lys (2) 0.1
500μM (3) 27.4
Lys (3) 24.4
5mM (4) 52.1
Lys (4) 51.3
50mM (5) 77.3
Lys (5) 77.0
Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Alicia_Rasines_Mazo/Notebook/CHEM-571_Experimental_Biological_Chemistry/2014/10/28&oldid=839904"