Creating Protein Prep Solutions: What Ingredients Do You Need?

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Discussion Overview

The discussion revolves around the preparation of protein solutions, specifically focusing on the ingredients and methods for creating solutions containing tributylphosphine, SDS, and urea. Participants explore the necessary concentrations and solvents for these solutions.

Discussion Character

  • Technical explanation
  • Conceptual clarification
  • Debate/contested

Main Points Raised

  • One participant requests recipes for making protein prep solutions, specifically mentioning 1 mM tributylphosphine, 1% SDS, and 10 M urea.
  • Another participant explains the calculation of weights needed for 1 mM and 10 M solutions, noting that urea is solid while tributylphosphine is typically liquid.
  • A participant inquires about the appropriate solvent for dissolving SDS, questioning whether to use sterile water or buffers.
  • It is suggested that SDS and urea can be diluted in distilled deionized water, with a note that buffer requirements should be specified in the solution's name.
  • One participant mentions that tributylphosphine is stored in N-methyl-2-pyrrolidine according to a reference from Sigma.
  • Another participant advises checking the spec sheet for solubility information and contacting the manufacturer if the spec sheet is unavailable.
  • A later reply emphasizes the use of nanopure water for protein chemistry and reiterates that SDS concentration is typically expressed as w/v.

Areas of Agreement / Disagreement

Participants express varying opinions on the appropriate solvents for the solutions and the specifics of preparing them, indicating that multiple views remain on these topics.

Contextual Notes

Some assumptions about the solubility of the chemicals and the definitions of concentration types (w/v) are not fully resolved, and there are references to specific products that may not be universally applicable.

sotellme
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I have searched and searched till my eyes fall out, but can not find any recipes for making these solutions. Anyone can help me out?

1mM tributylphosphine
1%SDS
10 M Urea
 
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It mostly a matter of you calculating the weight required for 1 mM solution and a 10 M solution. Urea is solid and Tributylphosphine is usually in liquid. Both should be found in your lab. It is basic chemistry.

As far the SDS solution goes, it is probably w/v but it might be specifiy. So when dealing with percentages in a recipe in term of w/v, it is about an amount of grams per 100 mL of solution. Therefore, it would be 1 g of SDS per 100 mL of solution.
 
hI iAN,

I am thinking of the solvent (?) to dissolve these stuffs. For example for SDS solution, should i use sterile water or buffers?
 
SDS and Urea can be diluted into distilled deionized water. Usually, if a chemical solution requires to be prepare in a buffer, it should be mention in the name of solution.

For tributylphosphine, I am not sure but sigma keeps their 200 mM solution in N-methyl-2-pyrrolidine.
http://www.sigmaaldrich.com/catalog/search/ProductDetail/SIGMA/T7567

You may want to check this reference given by sigma.
Herbert, B., et al., Reduction and alkylation of proteins in preparation of two dimensional map analysis: why, when, and how?. Electrophoresis 22, 2046-57, (2001)
 
Whenever you're in doubt, check the spec sheet that comes with the chemicals for information on solubility. If the chemicals didn't come with a spec sheet and you aren't sure, contact the manufacturer for solubility information.
 
if you are doing protein chemistry you should really use nanopure water.

% sds is almost always w/v
 

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