From now on you can become a member of this channel! You will have access to some cool emojis like this: . This is of course voluntary, but the financial support helps me a lot.
Why do we need two antibodies? Is it not possible to adjust the first antibody into one that can be conjugated by the peroxidase and is complementary towards the protein?
In some cases, there are primary antibodies that are already conjugated with a detection label, but these are usually limited to a few specific types of labels, and the specificity and sensitivity of these conjugated antibodies can sometimes be lower compared to using a secondary antibody. The use of two different antibodies in western blot is to increase the specificity of protein detection. This two-step process ensures that the signal generated is specific to the target protein, rather than non-specific binding of the detection reagents. It is possible to change the first antibody into one that can hold the marker, but it is highly laborious to do so. The process of conjugating a detection label to an antibody requires specialized expertise and conditions, which can be difficult to replicate for a large number of antibodies.
Thanks. But you should draw proteins in line shape, not in folding shape.because after SDS PAGE, all folding shapes of proteins change into line shape as we use Beta-mercaptoethanole.
Thank you. Yours is the only really useful channel on YT for biochemical methods I could find. As final revision for my exams these videos are perfect - all the details pop into my mind thanks to you so clearly recapitulating the basics for me.
@@henrikslab I mean isn't it the opposite (the membrane is placed on top of the gel) ? because the gel now contains the migrated proteins and we need to keep that in place. If I give you some pudding on a table and told you to lift that pudding and put it on top of a membrane without making a mess, that's hard..
@@yeseenmeshleeah7359 I did it several times my own, you have to be careful, but in the end it is not super hard. And there are many different ways (and clever tricks) to place the gel. The word "pudding" was a metaphor, most times it is more solid
@@yeseenmeshleeah7359 Like I said, there are so many cool hacks to do that best.. one is indeed, to place the membrane on top of the gel and then turn it together.. It is just extremely important to always make sure that they are placed in the right order (The electric current has to draw the proteins from gel onto membrane).
This is not true! Indeed the cathode for western blot is negatively charged (-) and the anode is positively charged (+) here. It is a common confusion for students, and I remember me asking the same question in one course... (the confusion comes from the fact that cations are positively charged, right?) Best Henrik
From now on you can become a member of this channel! You will have access to some cool emojis like this: .
This is of course voluntary, but the financial support helps me a lot.
Explained this better than my 4 hour lab. Thank you
great explanation! thank you so much, now i understand western blotting!
Such an excellent explanation
You helped me understand it within 4 minutes. Thanks 👍
Wow.... Hats off to your explanation🔥🔥🔥🔥
This was so clear to understand! Thank you so very much!
Thanks very helpful
this is so helpful thanks !
Thank you!
Why do we need two antibodies? Is it not possible to adjust the first antibody into one that can be conjugated by the peroxidase and is complementary towards the protein?
In some cases, there are primary antibodies that are already conjugated with a detection label, but these are usually limited to a few specific types of labels, and the specificity and sensitivity of these conjugated antibodies can sometimes be lower compared to using a secondary antibody.
The use of two different antibodies in western blot is to increase the specificity of protein detection. This two-step process ensures that the signal generated is specific to the target protein, rather than non-specific binding of the detection reagents. It is possible to change the first antibody into one that can hold the marker, but it is highly laborious to do so. The process of conjugating a detection label to an antibody requires specialized expertise and conditions, which can be difficult to replicate for a large number of antibodies.
Good work
Love ya
Is six weeks definitive for this analysis hiv
Shukriya
Hi Henrik! This technique seems strikingly similar to an enzyme-linked immunoassay test. Is it non the same?
Thanks. But you should draw proteins in line shape, not in folding shape.because after SDS PAGE, all folding shapes of proteins change into line shape as we use Beta-mercaptoethanole.
Thank you Henrik, brilliantly explained and simplified, keep them coming please
THANKS! You explaned the most important parts so well.
Thank you. Yours is the only really useful channel on YT for biochemical methods I could find. As final revision for my exams these videos are perfect - all the details pop into my mind thanks to you so clearly recapitulating the basics for me.
Anyone here because of MCAT prep? oooh well, maybe just me
IT´s brilliant how you make something looks so complicated in a very simple way i love ur vidos and ur talent
The animations really help a lot to understand the concept! Thank you!!
Thank you for good vedeo. I can development
I apologize but isn't the anode negatively charged?
No dude thats anions! Anions are negatively charged particles that move towards anode which is positively charged :D
Why can't they just have an enzyme limked to the first antibody, as with the way they'd carry out direct ELISA?
Explained than my professor. This is legit.
Thank you so much!!
Henrik, ich küsse dein Herz. Du rettest mich bei meinem Biochemie-Praktikum! Danke!
Medizin?
@@ornulusoundeffects6423Ja 😁
Schön visuell und super verständlich erklärt! Danke dir
Thank you.. so much
Bro if i could give you a hug🙂
You saved me for my exams 💜
Wow! Amazingly explained, very easy to understand! Thank you so much for this.
Many thanks
Thank you so much! I felt very lost in my lab prep but this makes so much sense now :)
Omg so helpful!! Thanks for making this animation!!!
Excelente video, gracias :)
Die videos auf deutsch wären nett :)
Brilliant!
this the best course in my life Thank you habibi!
Behind helpful. Thank you.
Excellent explanation, so glad I found this video.
Happy to hear!
❤️❤️❤️❤️
Very well-explained, Thanks!!!!!
Bro you are frkn amazing god bless you
amazing! i got all the info for my thesis. thank u
A ver isto pra aula de bcm da nova
Thank you for making me understand Western blot steps concise and clear.
Thank you so much!
Great explanation! thank you
Thank you so much
phenomenal explanation! thank you !
thanks ,God please you
thank you henriukkkk
thank you kind sir
Best science channel on youtube!
This was so helpful, thank you!!!
Truly a great video!
讚
Well explained thanks!
Thank you very much 🤩
Thanks you
perfect. thank you.
short and sweet! thank u!
Thank you Henrik!
LEER LOS ARTICULOS después de ver tus videos jajaja, es ley.
Very useful
capo, gracias!
Very good video!
Thanks
can you make a video on isoelectric foccusing please
Not planned at the moment, but I put it onto the long list of ideas!
ajudou muito, vlw🙃
Good meme
you just save me
great video
is this the principle for luminex as well?
Great clear explanation
Amazing
thanks! now I finally get it
Excellent and amazingly concise explanation! Thank you!
Great, thanks, but how is the gel (a liquid !) placed on top of the membrane , in practice ?
The gel (in the traditional blots) is solid.
It has the consistency of a pudding!
@@henrikslab I mean isn't it the opposite (the membrane is placed on top of the gel) ? because the gel now contains the migrated proteins and we need to keep that in place. If I give you some pudding on a table and told you to lift that pudding and put it on top of a membrane without making a mess, that's hard..
@@yeseenmeshleeah7359 I did it several times my own, you have to be careful, but in the end it is not super hard. And there are many different ways (and clever tricks) to place the gel. The word "pudding" was a metaphor, most times it is more solid
@@henrikslab Thank you so much for your reply. Your channel is a real marvel. Why not simply put the membrane on top ? Isn't it easier ?
@@yeseenmeshleeah7359 Like I said, there are so many cool hacks to do that best.. one is indeed, to place the membrane on top of the gel and then turn it together.. It is just extremely important to always make sure that they are placed in the right order (The electric current has to draw the proteins from gel onto membrane).
Brilliantly explained. Thank you 😊
that was so helpful☺ thank you so much
short. simple. easy to understand. tysm
Thnk u
You´re welcome, Kakashi
very simple and amazing, thanks alot
it is a helpful video!!
Thanks a lot.
There is a little mistake The Cathode is + and the Anode is -
apart from that I gave you a Like (The video is perfectly explained) Thank you very much for having posted it!
This is not true!
Indeed the cathode for western blot is negatively charged (-) and the anode is positively charged (+) here. It is a common confusion for students, and I remember me asking the same question in one course... (the confusion comes from the fact that cations are positively charged, right?)
Best
Henrik
@@henrikslab I apologize you are right ! Thank you Henrik!
@@MrWhite-fm4lj No need for that... it is a classical confusion!
@@henrikslab 😄😉
Very useful! thank you!
Thanks a lot 🤗