Mixture of proteins examine by standard 2-d electrophoresis

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Suppose you are given a mixture of proteins that you analyze by standard 2-D electrophoresis, with the following results for isoelectric points and apparent molecular weights: protein A (Mr 121,201; pI 8.22), protein B (Mr 55,438; pI 7.44), and protein C (Mr54,274; pI 10.54).

  1. With the SDS PAGE, in what order did the proteins band, starting with the well and proceeding to the bottom of the gel? Explain why.

  2. If the mixture is subjected to CM-matrix chromatography, what pH range should you use for the buffer and in what order will the proteins emerge from the column? Explain why.

  3. Upon gel filtration of the mixture, protein A emerges from a G2000 Sephadex column at a Ve of 115 ml protein B emerges at 155 and protein C at 190 ml. Based on this observation and the 2-D electrophoresis results, what can you conclude about the structure of protein C? Explain.

  4. Upon salting out with ammonium sulfate, if you use pH 7.0 for the buffer, in what order should the proteins precipitate? Explain

Reference no: EM1383133

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