User:Michael S. Bible/Notebook/571/2014/09/16: Difference between revisions
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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span> | |style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</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}}}}]]}} | ||
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==Description== | ==Description== | ||
*Today we dialyzed a 35:1 Au:Protein solution (colloid solution) made earlier and stored in our lab drawer. | |||
* | *For dialysis, we needed to create 1 L of a 50 mM Glycine buffer, with a pH ~3.5. | ||
**To do this, 4 mL of 1 M HCl and 3.7608 grams of glycine were combined in a 1 L volumetric flask. | |||
*We also needed to prepare 500 mL of a 50 mM NaCl solution. | |||
**To do this, 1.46307 g of NaCl was added to water in a 500 mL volumetric flask. | |||
*Additionally, 25 mL of 1 g/L Lysozyme was prepared. | |||
**To do this, 0.025 g of Lysozyme was combined with water in a 25 mL volumetric flask. | |||
*We then performed dialysis using the protocol described in the "Dialysis Protocol" section below to prepare the dialysis tubes. | |||
**Dialysis was performed by adding 4 mL of the lysozyme solution to: | |||
***A 3500 g/mol MWCO tube in NaCl soak. | |||
***A 3500 g/mol MWCO tube in Glycine Buffer soak. | |||
***A 3500 g/mol MWCO tube in HPLC water soak. | |||
***A 25000 g/mol MWCO tube in Glycine Buffer soak. | |||
**An additional dialysis was performed using 4 mL of the colloid solution in: | |||
***A 3500 g/mol MWCO tube in Glycine Buffer soak. | |||
==Dialysis Protocol== | |||
#Cut about a 3" length of dialysis tubing | |||
#*25,000 MWCO tubing is stored in an azide solution to prevent mold (use care; NaN<sub>3</sub> is toxic) | |||
#*keep the 25,000 MWCO tubing wet to prevent pore shrinkage | |||
#wash cut tubing with DI water, both inside and out | |||
#*3,500 MWCO and 15,000 MWCO are dry and need to be wet prior to filling | |||
#*25,000 MWCO needs to be rinsed to remove the sodium azide | |||
#flatten tubing and remove as much residual water as possible with gloves fingers | |||
#clamp one end 1/4 - 1/2" from edge | |||
#open other end and transfer your '''measured''' protein solution inside | |||
#carefully flatten open end and clamp, making sure no liquid escapes | |||
#rinse with DI water, particularly the ends, which may have residual protein solution on it | |||
#place into a 150 mL beaker (or 250 mL or 400 mL) | |||
#measure out 100 - 200 mL of your dialyzing solution | |||
#label beaker, cover with parafilm, and leave on shaker on low speed to help prevent gradients during dialysis | |||
[[Category:Course]] | [[Category:Course]] |
Latest revision as of 00:18, 27 September 2017
Biomaterials Design Lab | Main project page Previous entry Next entry |
Dialysis of Colloid SampleProtocols for today were adopted from Dr. Fox's Notebook. Description
Dialysis Protocol
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