Why do we need all three salts in MOPS buffer? and PEG added to the reagent.
NaCl crystalline 1.359 g/L
KCl 0.708 g/L
Ammonium chloride 0.0063 g/L
KCL might precipitate the protein PEG and NH4CL solubilize it?
MOPS (3-(N-Morpholino)propanesulfonic acid) is a buffer substance required to adjust the pH, together with some acid that protonates the ring nitrogen (the sulphonic acid group is a strong acid and fully deprotonated at physiological pH).
NaCl (23.25 mM), KCl (9.5 mM) and NH4Cl (0.12 mM) are probably used to mimic the ionic environment, but without info on what the buffer is used for it is impossible to be more specific.
PEG (polyethylene glycol) is used as osmolyte, it reduces the active water concentration and hence prevents intra-molecular movement of proteins. This reduces the enzymatic activity, increases stability and makes proteins easier to crystallise. It may also adjust the osmotic pressure of the solution. Note that usually there is a number (several hundred to several thousand) behind the abbreviation PEG which denotes the average Mr.
ProClin 300 is a thiazolin-based preservative that inhibits dehydrogenases, and hence the Krebs-cycle. Be careful with it, it is not only toxic but a known contact allergen.
BSA is commonly used to protect other proteins against unspecific interactions (aggregation), both amongst themselves and with the vial wall.
MOPS (3-(N-Morpholino)propanesulfonic acid) is a buffer substance required to adjust the pH, together with some acid that protonates the ring nitrogen (the sulphonic acid group is a strong acid and fully deprotonated at physiological pH).
NaCl (23.25 mM), KCl (9.5 mM) and NH4Cl (0.12 mM) are probably used to mimic the ionic environment, but without info on what the buffer is used for it is impossible to be more specific.
PEG (polyethylene glycol) is used as osmolyte, it reduces the active water concentration and hence prevents intra-molecular movement of proteins. This reduces the enzymatic activity, increases stability and makes proteins easier to crystallise. It may also adjust the osmotic pressure of the solution. Note that usually there is a number (several hundred to several thousand) behind the abbreviation PEG which denotes the average Mr.
ProClin 300 is a thiazolin-based preservative that inhibits dehydrogenases, and hence the Krebs-cycle. Be careful with it, it is not only toxic but a known contact allergen.
BSA is commonly used to protect other proteins against unspecific interactions (aggregation), both amongst themselves and with the vial wall.
Well, How does NACL affect the specificity of PCR? Does it indeed cause an increase or decrease?
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