16NOV17 purification small scale IRM12
16NOV17
I prepared some more buffers (I used 2.5% glycerol this time just to reduce the glycerol a bit since it blocks the filters and the protein is defficult to be concentrated with the cut off filter tubes) for IRM12 and purified it. I run the gels for both IRM12 and IRM13.
I could see IRM13 in the last 2 elution fractions (250mM and 350mM Imidazole) but IRM12 was no visible. So I continue with dialysing the IRM13.
I made 1.5 lt of TRIS 50mM buffer with 150mM NaCl and pH adjusted in a 2lt beaker. I put the 2 elution fractions in a snakeskin bag and put it in the beaker and left it opvernight on a stirrer with a stirring stone in the cold room.
I prepared some more buffers (I used 2.5% glycerol this time just to reduce the glycerol a bit since it blocks the filters and the protein is defficult to be concentrated with the cut off filter tubes) for IRM12 and purified it. I run the gels for both IRM12 and IRM13.
I could see IRM13 in the last 2 elution fractions (250mM and 350mM Imidazole) but IRM12 was no visible. So I continue with dialysing the IRM13.
I made 1.5 lt of TRIS 50mM buffer with 150mM NaCl and pH adjusted in a 2lt beaker. I put the 2 elution fractions in a snakeskin bag and put it in the beaker and left it opvernight on a stirrer with a stirring stone in the cold room.
Comments
Post a Comment