Creating Protein Prep Solutions: What Ingredients Do You Need?

[sotellme]

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

 

[iansmith]

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.

 

[sotellme]

hI iAN,

I am thinking of the solvent (?) to dissolve these stuffs. For example for SDS solution, should i use sterile water or buffers?

 

[iansmith]

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)

 

[Moonbear]

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.

 

[quetzalcoatl9]

if you are doing protein chemistry you should really use nanopure water.

% sds is almost always w/v