how do we separate the modified glutamine and Met.Lys. with the protein, in case we only want that pure protein as a result? can someone help me please thank you so much
What? How can histidine tagged enzymes be in the same column as the POI if they cleave the histidine tags from them? Wouldn't the POI get eluted out as soon as it is put into the column?
Insomniac it’s a two step column purification. First, the his-tagged protein of interest is bound and eluted, purified from other host cell proteins. Then the his tagged enzymes are added to eluate, cleaving the POI’s his tag. The mixture of his tagged enzymes and POI (with his tagged cleaved) are loaded onto a nickel column a second time (after removing imidazole of course), and the enzymes bind to the column while the POI is in the flow through this time.
![I.N "HALLUCINATION" | [Stray Kids : SKZ-PLAYER]](http://i.ytimg.com/vi/n5B5q1Hwt_U/mqdefault.jpg)
Relatively quickly "IN THE PERIOD OF A WEEK"
IT TAKES FOREVER!!!!!!!!! LIKE 2 DAYS STRAIGHT NONSTOP
AMAZINGLY HELPFUL AND SIMPLE!!!!!
it's better video for easy understanding abot his tag.... thnq you
So clearly thx so much im gonna test about this in the next few hours😂😂😂
fog windd how did you do
Very good video!!
Great explanation
Great animation. Thanks :)
THANK YOU SO MUCH
thank you!!
how do we separate the modified glutamine and Met.Lys. with the protein, in case we only want that pure protein as a result? can someone help me please thank you so much
i guess
we can use specific proteases
Awesome
What? How can histidine tagged enzymes be in the same column as the POI if they cleave the histidine tags from them? Wouldn't the POI get eluted out as soon as it is put into the column?
Insomniac it’s a two step column purification. First, the his-tagged protein of interest is bound and eluted, purified from other host cell proteins. Then the his tagged enzymes are added to eluate, cleaving the POI’s his tag. The mixture of his tagged enzymes and POI (with his tagged cleaved) are loaded onto a nickel column a second time (after removing imidazole of course), and the enzymes bind to the column while the POI is in the flow through this time.
Want biotechnology video how I get it
what are the enzymes used for cutting the 6xHis tags on the proteins
you still need that information bro ?
@@wadjo1 I'm curious to know
@@wadjo1 yep
@@wadjo1 I need it
Just subscribed, keep it goin. I'm trying to grow my RUclips channel too, it's hard but it's still fun. Peace!
ugh, i hate the synthetic voice thing.. sorry.
imi-DAZ-ol lol