https://bionmr.unl.edu/mediawiki/mediawiki/index.php?action=history&feed=atom&title=Digestion_Recipies 2026-05-02T13:54:01Z Revision history for this page on the wiki MediaWiki 1.38.2 https://bionmr.unl.edu/mediawiki/mediawiki/index.php?title=Digestion_Recipies&diff=1300&oldid=prev 2022-04-22T22:31:06Z

<p>Created page with &quot;<a href="/mediawiki/index.php/Category:CIBC" title="Category:CIBC">category:CIBC</a> [https://uofnelincoln.sharepoint.com/:w:/r/sites/UNLChemistryDepartment/CIBC/Protocols/Proteomic%20Protocols/Digestion%20Recipes.docx?d=wfe52b1816ee549258ef...&quot;</p> <p><b>New page</b></p><div>[[category:CIBC]]<br /> <br /> [https://uofnelincoln.sharepoint.com/:w:/r/sites/UNLChemistryDepartment/CIBC/Protocols/Proteomic%20Protocols/Digestion%20Recipes.docx?d=wfe52b1816ee549258efce68eb68c440a&amp;csf=1&amp;web=1&amp;e=2PZy3Q Sharepoint File Link]<br /> <br /> ===Suggested In-Gel Digestion Solutions=== <br /> <br /> ⦁ AmBic 100 mM: Dissolve 1.40 g in 150 mL ultrapure H2O. Fill to 200 mL with H2O in volumetric flask. <br /> <br /> ⦁ AmBic 50 mM: Prepare from 100 mM AmBic solution with ultrapure H2O <br /> <br /> ⦁ Destain solution: Prepare fresh by mixing ACN with 100 mM AmBic (1:1, vol/vol). <br /> <br /> ⦁ Reduction solution: 10 mM DTT in 100 mM AmBic. Make shortly before use. <br /> <br /> ⦁ Alkylation solution: 55 mM IAA in 100 mM AmBic. Make shortly before use. IAA is light sensitive! Keep it in dark. <br /> <br /> ⦁ Trypsin stock solution 100 ng/µL: Add 200 µL 50 mM Acetic acid into a 20 µg trypsin vial. Mix by pipetting up and down in tip. Do not vortex! Store 30 µL aliquots at -80°C. <br /> <br /> ⦁ Trypsin working solution 15 ng/µL: Thaw one tube of stock solution on ice and add 170 µL of 50 mM AmBic. <br /> <br /> ⦁ Stop solution: 5% (v:v) formic acid in 50% (v:v) ACN. <br /> <br /> ⦁ Elution solution A: 1% (v:v) formic acid in 60% (v:v) MeOH. <br /> <br /> ⦁ Elution solution B: 1% (v:v) formic acid in 50% (v:v) ACN. <br /> <br /> ⦁ Wetting solution I: 100% LC-MS grade acetonitrile (ACN) <br /> <br /> ⦁ Wetting solution II: 50% LC-MS grade ACN <br /> <br /> ⦁ Equilibration &amp; Sample solutions: 0.1% TFA in HPLC water <br /> <br /> ⦁ Washing solution: 0.1% TFA in 5% LC-MS grade Methanol <br /> <br /> ⦁ Elution solution: 0.1% TFA in 50% ACN</div>

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