thank you. here because my proteins transferred right into my buffer. i didn’t realize what i’d done wrong until you pointed out the nitrocellulose membrane needed to face the clear side of the cassette.
We place the whole cassette into the casserole dish, making sure it is already in the correct orientation and build the sandwich right in the cassette. The casserole dish has tranfer buffer from the previous run. Place the first sponge in the cassette and the second can just sit in the dish along side. Then place one filter paper on top of each sponge. Finally, place the nitrocellulose on the filter paper in the cassette. Add the gel, cover with paper then sponge. Close cassette
The lady in the video put 2 NC membranes on top of a blot paper then put the gel containing the proteins on top of the NC membranes. Now I know that is fine, however she tried to press the gel on the NC membranes to avoid bubbles. I am not sure that is quite efficient because (1) it's hard to see the bubbles on the gel is on top and (2) why did she have to flip the gel? The proteins are actually stuck inside the gel not on one side, so it shouldn't matter what side the gel is.
![[Western Blot] 웨스턴 블롯 전반적인 과정 설명 : 기본편](/img/1.gif)
thank you. here because my proteins transferred right into my buffer. i didn’t realize what i’d done wrong until you pointed out the nitrocellulose membrane needed to face the clear side of the cassette.
Thanks for sharing this!! This was EXTREMELY helpful to a graduate student like myself!!! I appreciate you!!
We place the whole cassette into the casserole dish, making sure it is already in the correct orientation and build the sandwich right in the cassette. The casserole dish has tranfer buffer from the previous run. Place the first sponge in the cassette and the second can just sit in the dish along side. Then place one filter paper on top of each sponge. Finally, place the nitrocellulose on the filter paper in the cassette. Add the gel, cover with paper then sponge. Close cassette
The lady in the video put 2 NC membranes on top of a blot paper then put the gel containing the proteins on top of the NC membranes. Now I know that is fine, however she tried to press the gel on the NC membranes to avoid bubbles. I am not sure that is quite efficient because (1) it's hard to see the bubbles on the gel is on top and (2) why did she have to flip the gel? The proteins are actually stuck inside the gel not on one side, so it shouldn't matter what side the gel is.
Thank you so much for the help!!!!!!!!!!!!!!!!!!!
Thank you so much for the help.
Thank you for sharing this
so good
naitrocellyoulus