User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/29: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Autocreate 2014/10/29 Entry for User:Alicia_Rasines_Mazo/Notebook/CHEM-571_Experimental_Biological_Chemistry) |
(fix raw html notebook nav) |
||
(10 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"| | |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]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
==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 | |||
***0.5v<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
Preparation of HCl stock solutions
Preparation of new 0.6g/L lysozyme stock
Dialysis of 0.6 g/L Lysozyme vs HCl using MWCO 20000
Bradford Analysis
pH measurements
|