gst fusion protein
Posted 25 September 2010 - 11:47 PM
My question is: can i do gst pulldown even if i dont see induction from 1 ml of broth i took for coomassie blue staining? Can my gst-fusion be not strong enough to be detected in coomassie blue?
any of your advise will be helpful... thanks so much...
Posted 25 September 2010 - 11:53 PM
However, when i did gst purification (figure 2) , i see 4 bands in my gst-fusion sample. lane 1 is marker, lane 2 is gst purified, lane 3 is the gst-fusion bacterial sample.
grateful for your advise.
Posted 27 September 2010 - 04:51 PM
Also, even though you have something that appears to be the right size bound to your beads, you also have several possible breakdown products, which is not optimal. Ideally you don't want any breakdown products. Either way your protein should definitely be in excess of the breakdown products. Are you using protease inhibitors?
Posted 28 September 2010 - 04:46 PM
yes, i used protease inhibitors to lyse and sonicate my bacterial pellets. i am grateful for your answer and for spending time taking a look at this. i hope i get better induction soon....
if there are any advise, tips or comments you may suggest, i will be very happy to learn from you all. thank you so much....
Edited by soymilk14, 28 September 2010 - 04:48 PM.