Western Blot / Protein Immunoblot explained

Hey Friends,
Western Blot is a technique to determine whether a specific protein is present in a sample. Before, the proteins were separated according to their Molecular Weight using Polyacrylamide Gel Electrophoresis.
Short video about SDS-PAGE:
• SDS-PAGE explained - P...
Western Blot allows to transfer the proteins from that gel onto a membrane. This membrane is then incubated with antibodies to detect the presence of the protein of desire.
Very cool sheet about Western Blot:
precisionbiosystems.com/weste...
Thanks for watching!
Henrik
Chapters:
0:00 Introduction
0:22 SDS PAGE
0:47 Gel after SDS PAGE
1:25 Western Blot
4:12 Outro

[@henrikslab]

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.

[@Grngrsky]

Explained this better than my 4 hour lab. Thank you

[@ilm15]

Henrik, ich küsse dein Herz. Du rettest mich bei meinem Biochemie-Praktikum! Danke!

[@mahyammm]

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.

[@doktorsugar1609]

Thank you Henrik, brilliantly explained and simplified, keep them coming please

[@user-oh6be6fm5m]

The animations really help a lot to understand the concept! Thank you!!

[@rickinho99h]

Excellent and amazingly concise explanation! Thank you!

[@juditaanthi9284]

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.

[@Mia_xingyu]

THANKS! You explaned the most important parts so well.

[@cedrictorndahl5945]

Wow! Amazingly explained, very easy to understand! Thank you so much for this.

[@giddygiddybangbang9425]

Thank you for making me understand Western blot steps concise and clear.

[@sofiar5513]

Schön visuell und super verständlich erklärt! Danke dir

[@dimahalshehnah4199]

IT´s brilliant how you make something looks so complicated in a very simple way i love ur vidos and ur talent

[@farhanahassan8795]

Brilliantly explained. Thank you 😊

[@kalkidanberhanu7708]

Thank you so much! I felt very lost in my lab prep but this makes so much sense now :)

[@andersmannelqvist1157]

Thank you Henrik!

[@kagamer21]

This was so clear to understand! Thank you so very much!

[@klam77]

phenomenal explanation! thank you !

[@nassimabk3714]

short and sweet! thank u!

[@kamrunnaharshefa8471]

You helped me understand it within 4 minutes. Thanks 👍

[@ahmetziyaaktas9179]

Such an excellent explanation

[@ibqvelazquezlae]

Very well-explained, Thanks!!!!!

[@maestros1327]

this the best course in my life Thank you habibi!

[@NoopyP]

This was so helpful, thank you!!!

[@eeshaumer607]

short. simple. easy to understand. tysm

[@diellzadibrani6122]

Truly a great video!

[@guiyeoullim1778]

Great explanation! thank you

[@shreyaskulkarni5312]

Wow.... Hats off to your explanation🔥🔥🔥🔥

[@shaylanmoodley7056]

Brilliant!

[@merveggundogdu]

amazing! i got all the info for my thesis. thank u

[@paulinahiyaev9712]

Thank you so much!

[@kristianpelligra2588]

Excellent explanation, so glad I found this video.

[@henrikslab]

Happy to hear!

[@saiman7075]

Best science channel on youtube!

[@palvoliuto6059]

great explanation! thank you so much, now i understand western blotting!

[@sarthakthememegod]

Thank you so much!!

[@amiramohamed4880]

very simple and amazing, thanks alot

[@dobby0960]

thank you henriukkkk

[@kanchanbhamare899]

Well explained thanks!

[@lauraconceicao2300]

this is so helpful thanks !

[@vladpetre2873]

Very good video!

[@yasmeenmohamad4515]

that was so helpful☺ thank you so much

[@afsaneemami580]

perfect. thank you.

[@lindagao8171]

Omg so helpful!! Thanks for making this animation!!!

[@mohamedshembesh8688]

Bro you are frkn amazing god bless you

[@justanotheremptychannel8851]

thanks! now I finally get it

[@DavidAbisaics]

Excelente video, gracias :)

[@sarah-sn1ov]

Thank you very much 🤩

[@afsanesoltani7709]

Many thanks

[@nidamumtaz557]

Thanks very helpful

[@osmirod1]

Behind helpful. Thank you.

[@wangleeleiting2689]

great video

[@biovalor]

Thank you!

[@sajibbanik5319]

Good work

[@GRACE-ii5vf]

Thank you.. so much

[@antonionave7247]

Thank you so much

[@jayashreekbiotech9995]

Very useful

[@tanmoyadhikari1851]

Bro if i could give you a hug🙂
You saved me for my exams 💜

[@snarki1906]

thank you kind sir

[@60SekundenBucher]

Thanks a lot.

[@user-sd8vn3tn4g]

Amazing

[@fabiolarusso7023]

Very useful! thank you!

[@gusttk]

is this the principle for luminex as well?
Great clear explanation

[@rachmiputri]

it is a helpful video!!

[@tarteelmohammed3257]

Thanks you

[@camilabrazuna3841]

capo, gracias!

[@TANMAN47TANMAN]

Thanks

[@Tepmodify]

Thank you for good vedeo. I can development

[@luanatifany4361]

ajudou muito, vlw🙃

[@khyatipandey3734]

Thanks a lot 🤗

[@phmarwa8616]

thanks ,God please you

[@Pattii248]

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?

[@henrikslab]

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.

[@bestking5727]

Is six weeks definitive for this analysis hiv

[@user-sk4dp2pk5n]

can you make a video on isoelectric foccusing please

[@henrikslab]

Not planned at the moment, but I put it onto the long list of ideas!

[@freedomtv1505]

Love ya

[@rajeshbal6592]

Thnk u

[@henrikslab]

You´re welcome, Kakashi

[@tayyabaliqaiser442]

Shukriya

[@yeseenmeshleeah7359]

Great, thanks, but how is the gel (a liquid !) placed on top of the membrane , in practice ?

[@henrikslab]

The gel (in the traditional blots) is solid.
It has the consistency of a pudding!

[@yeseenmeshleeah7359]

@@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..

[@henrikslab]

@@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]

@@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 ?

[@henrikslab]

@@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).

[@pitkamatka]

Hi Henrik! This technique seems strikingly similar to an enzyme-linked immunoassay test. Is it non the same?

[@muhindoedith5459]

❤️❤️❤️❤️

[@danwittek985]

I apologize but isn't the anode negatively charged?

[@lee7915]

讚

[@ramapozaodwyer6317]

Anyone here because of MCAT prep? oooh well, maybe just me

[@Raven-hz8ou]

you just save me

[@ferricyanide8112]

Good meme

[@MrWhite-fm4lj]

There is a little mistake The Cathode is + and the Anode is -

[@MrWhite-fm4lj]

apart from that I gave you a Like (The video is perfectly explained) Thank you very much for having posted it!

[@henrikslab]

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

[@MrWhite-fm4lj]

@@henrikslab I apologize you are right ! Thank you Henrik!

[@henrikslab]

@@MrWhite-fm4lj No need for that... it is a classical confusion!

[@MrWhite-fm4lj]

@@henrikslab 😄😉

[@alfredoperez4595]

LEER LOS ARTICULOS después de ver tus videos jajaja, es ley.

[@Aio9818]

Die videos auf deutsch wären nett :)

[@mak5853]

Thanks