All those collecting tubes should be placed on ice. Also, cell lysis was not shown, and protein extraction from lysate, before Nickel bead affinity chromatography was not even shown, but i guess whatever...
Hi Rebecca, thank you. I am curious how one can know how much wash to use if you are purifying a protein that cannot be seen like GFP? Is one column of wash good enough usually and then you can use the imidazole eluent? Thank you.
Hey Rebecca, Is that so called "cloudiness" a Biofilm that the Bacteria had produced ? If not, then what is it ? There has got to be a faster way to remove the GFP. lol Thank-You.
I loved the spooky sound in between 😂😂😂😂
so do I
Would have been spooked myself had I not read your comment before starting to watch the video :-D
thanks rebeca
thank you, Rebecca.
Nice demo Rebecca!!
Great video, thank you
Great Demonstration ♥
very helpful. thanks.
i love this video
I think that the GFP if I remember correctly
Awesome!
Did Isaiah 46:9 say Jesus is God? Check your facts
thks so much
Thanks!!!
All those collecting tubes should be placed on ice. Also, cell lysis was not shown, and protein extraction from lysate, before Nickel bead affinity chromatography was not even shown, but i guess whatever...
Hi Rebecca, thank you. I am curious how one can know how much wash to use if you are purifying a protein that cannot be seen like GFP? Is one column of wash good enough usually and then you can use the imidazole eluent? Thank you.
Who can let me know how to determine agent for purification with curli in E Coli? :(
May I know what is the method that you used to break the cell? how pure is the final product?
I am not sure what she used, but often sonication is used to lyse the cell. That's what I use for my lab anyways.
yeah, sonication can be the answer or, for bigger volumes, you could use a French press (reverse system) method
no it's just pure biomass you are seeing, there are simply more bacteria than before.
At the end, the chemical used for removing gfp from beads needed to be separated from the protein. I think that could have been mentioned as well.
Ice job
Hi, how did you isolated the proteins from the e.coli? which method?
By IMAC chromatography.
In case you steel looking for the answer 7 years after you wrote it :)
Where is the dialysis?
sister which protein do you want to purify?
Just subscribed, keep it goin. I'm trying to grow my TH-cam channel too, it's hard but it's still fun. Peace!
gfp
you did not even show people on how to use the sonicator or even vortexing the pellet with buffer.
I love this girl, would you marry me? X-D!
Nice try bud. She is mine.
She will become a nun
i came here for protien purification but i dont care now , this girl s damn beautiful
thank you its easy way for purification,,
i need protocol for purify topoisomerase IV from pseudomonas aeruginosa.
can you guide me.
thank you
Do they have different polarities? If so maybe you could use column chromatography?
Hey Rebecca,
Is that so called "cloudiness" a Biofilm that the Bacteria had produced ? If not, then what is it ? There has got to be a faster way to remove the GFP. lol Thank-You.
It's the optical density/absorbance of the bacterial cells
how deleted my comments....... mo... f.....s!!, yeah I love this, so what, any problem? B-D!
Now I am happy again! thanks! :-D!