i have 4 lanes worth of samples. it is always 1 lane away from the protein markers. it is at lane 2, 5, 8, 9
the amount of total protein lysate is 60ug. the first image is 10 seconds after water destain, the 2nd image is about 1 minute water destaining.
is there any general degradation of proteins or abnormalities seen here?
I don't see what could be abnormal : the first image shows a sheet that is partially destained and the second image a fully destained one. As you will have noticed, the Ponceau red stain is not stable and will disappear of you destain longer. Thus itis advisable to take a picture of the membrane, or to mark with pencil the position of the bands of interest if you want to keep a permanent record.
Hi, I agree with the previuos answer, in general when I want to see a good Ponceau I stain with ponceau for few second with fresh solution, a little more with old or diluited (recycled) one. Wash very quickly with water anf take a photo avoiding to dry the membrane, then destain with PBS (better the water). You wash after the ponceau so the membrane begin to destain and you have an uneven pattern (maybe you used a ponceau recycled a lot of time). You have a big band, maybe a protein very high expressed that remain also after the destain, the others simple disappear faster. First try to leave more ponceau or use fresh one, photo before destain. There are not smear it is only the ponceau not stained well, not washed away well.
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