User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/29: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
(fix raw html notebook nav)
 
(7 intermediate revisions by one other user not shown)
Line 2: Line 2:
|-
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
|-
|-
| colspan="2"|
| colspan="2"|
==Tasks for October 29===
==Tasks for October 29==
*Dialysis of 0.6 g/L Lysozyme vs HCl using MWCO 20000
*Dialysis of 0.6 g/L Lysozyme vs HCl using MWCO 20000


Line 11: Line 11:
* Need to prepare HCl stock solution of 75 μM, 100 μM, 250 μM, 500 μM and 5 mM with distilled water.  
* Need to prepare HCl stock solution of 75 μM, 100 μM, 250 μM, 500 μM and 5 mM with distilled water.  
**The initial stock solution of 5 mM HCl was prepared by diluting 0.5 mL of 0.5 N HCl to 50 mL using distilled water
**The initial stock solution of 5 mM HCl was prepared by diluting 0.5 mL of 0.5 N HCl to 50 mL using distilled water
***0.5v<sub>1</sub>=5×10<sub>-3</sub>(50mL); '''v<sub>1</sub>=0.5 mL'''
***0.5v<sub>1</sub>=5×10<sub>-3</sub>(50mL); '''v<sub>1</sub>=0.5 mL.'''
**Made remaining stock solutions from 5 mM stock


===Preparation of new 0.6g/L lysozyme stock===
* Had to prepare new 0.6 g/L lysozyme since it precipitated out of solution.
* Made 50 mL of 0.6 g/L lysozyme solution by adding 0.0380 g of lysozyme (Theoretical value 0.030 g, but some may have been lost in transfer).
===Dialysis of 0.6 g/L Lysozyme vs HCl using MWCO 20000===
* '''Note''': 100 μM HCl completed evaporated from well, despite being screwed shut.
<u>Bradford Analysis</u>
*Ran bradford analysis of all protein-containing solutions, that is Lysozyme 1, 2, 3, 4 and 5, and all the HCl solutions.
* Remove 100 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, 100 μL solution + 200 μL diluted Bradford + 700 μL Tris/NaCl buffer
**PS cuvettes, measuring 400 - 800 nm
<u>pH measurements</u>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Substance'''
| align="center" style="background:#f0f0f0;"|'''pH measurement'''
|-
| 75 μM HCl (1) || 7.142
|-
| Lys 1 || 7.116
|-
| Lys 2 || 7.423
|-
| 250 μM HCl (3) || 5.707
|-
| Lys 3 || 6.158
|-
| 500μM HCl (4) || 4.944
|-
| Lys 4 || 4.921
|-
| 5mM HCl (5) || 2.996
|-
| Lys 5 || 3.025
|-
|}
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->

Latest revision as of 00:29, 27 September 2017

Project name Main project page
Previous entry      Next entry

Tasks for October 29

  • Dialysis of 0.6 g/L Lysozyme vs HCl using MWCO 20000

Preparation of HCl stock solutions

  • Need to prepare HCl stock solution of 75 μM, 100 μM, 250 μM, 500 μM and 5 mM with distilled water.
    • The initial stock solution of 5 mM HCl was prepared by diluting 0.5 mL of 0.5 N HCl to 50 mL using distilled water
      • 0.5v1=5×10-3(50mL); v1=0.5 mL.
    • Made remaining stock solutions from 5 mM stock

Preparation of new 0.6g/L lysozyme stock

  • Had to prepare new 0.6 g/L lysozyme since it precipitated out of solution.
  • Made 50 mL of 0.6 g/L lysozyme solution by adding 0.0380 g of lysozyme (Theoretical value 0.030 g, but some may have been lost in transfer).

Dialysis of 0.6 g/L Lysozyme vs HCl using MWCO 20000

  • Note: 100 μM HCl completed evaporated from well, despite being screwed shut.

Bradford Analysis

  • Ran bradford analysis of all protein-containing solutions, that is Lysozyme 1, 2, 3, 4 and 5, and all the HCl solutions.
  • Remove 100 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, 100 μL solution + 200 μL diluted Bradford + 700 μL Tris/NaCl buffer
    • PS cuvettes, measuring 400 - 800 nm

pH measurements

Substance pH measurement
75 μM HCl (1) 7.142
Lys 1 7.116
Lys 2 7.423
250 μM HCl (3) 5.707
Lys 3 6.158
500μM HCl (4) 4.944
Lys 4 4.921
5mM HCl (5) 2.996
Lys 5 3.025
Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Alicia_Rasines_Mazo/Notebook/CHEM-571_Experimental_Biological_Chemistry/2014/10/29&oldid=1015734"