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re-format and add non-bacterial
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Several methods are available for the bacterial expression of semet-labelled proteins.
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= bacterial expression of semet-labelled proteins =
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In the literature:
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references:
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400 ml M63 stock salts
 
400 ml M63 stock salts
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20 ml of [http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins#solution_1: solution 1]
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# 20 ml of solution 1
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20 ml of [http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins#solution_2: solution 2]
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for 50 ml (in ddH<sub>2</sub>O)
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add 200 mg of the following amino acids:
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A, R, G, Q, H, I, L, K, P, S, T, V
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pH should be adjusted to 7-7.5 with 1 M KOH
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# 20 ml of solution 2
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20 ml of [http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins#solution_3: solution 3]
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for 50 ml (in ddH<sub>2</sub>O)
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add 200 mg of the following amino acids:
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N, D, C, E
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pH should be adjusted to 7-7.5 with 1 M KOH
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20 ml of [http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins#solution_4: solution 4]
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# 20 ml of solution 3
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20 ml of [http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins#solution_5: solution 5]
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for 50 ml
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200 mg phenylalanine
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100 mg tryptophane
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200 mg tyrosine
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20 mg p-aminobenzoic acid
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20 mg p-hydrobenzoic acid
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dissolve in 50 ml 0.1 M KOH
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# 20 ml of solution 4
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for 50 ml
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200 mg hypoxanthine
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200 mg uracil
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dissolve in 50 ml DMSO
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# 20 ml of solution 5
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for 500 ml (in ddH<sub>2</sub>O)
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100 mg biotin
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100 mg nicotinamide
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10 mg riboflavin
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100 mg thiamine
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# M63 stock solution
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for 1 liter (in ddH<sub>2</sub>O)
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15 g KH<sub>2</sub>PO<sub>4</sub>
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45.6 g K<sub>2</sub>HPO<sub>4</sub>
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10 g (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>
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2.5 ml FeSO<sub>4</sub> at 2.5 mg/ml
    
2 ml of 1 M MgSO<sub>4</sub> stock
 
2 ml of 1 M MgSO<sub>4</sub> stock
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== M63 stock solution: ==
  −
  −
for 1 liter (in ddH<sub>2</sub>O)
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  −
15 g KH<sub>2</sub>PO<sub>4</sub>
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45.6 g K<sub>2</sub>HPO<sub>4</sub>
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10 g (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>
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2.5 ml FeSO<sub>4</sub> at 2.5 mg/ml
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  −
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== solution 1: ==
  −
  −
for 50 ml (in ddH<sub>2</sub>O)
  −
  −
add 200 mg of the following amino acids:
  −
A, R, G, Q, H, I, L, K, P, S, T, V
  −
  −
pH should be adjusted to 7-7.5 with 1 M KOH
  −
  −
== solution 2: ==
  −
  −
for 50 ml (in ddH<sub>2</sub>O)
  −
  −
add 200 mg of the following amino acids:
  −
N, D, C, E
  −
  −
pH should be adjusted to 7-7.5 with 1 M KOH
  −
  −
== solution 3: ==
  −
  −
for 50 ml
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200 mg phenylalanine
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100 mg tryptophane
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200 mg tyrosine
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20 mg p-aminobenzoic acid
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20 mg p-hydrobenzoic acid
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  −
dissolve in 50 ml 0.1 M KOH
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  −
== solution 4: ==
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  −
for 50 ml
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200 mg hypoxanthine
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200 mg uracil
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dissolve in 50 ml DMSO
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== solution 5:  ==
     −
for 500 ml (in ddH<sub>2</sub>O)
     −
100 mg biotin
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= expression of semet-labelled proteins in other systems =
100 mg nicotinamide
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* mammalian cells: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2242577/pdf/2008.pdf mentions several mammalian cell lines, including CHO, HEK239, and COS. Apparently, Lec8 have been used, too.
10 mg riboflavin
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* Yeast
100 mg thiamine
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* stable S2 insect cells
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* baculovirus expression system
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