Biotechniques | General Theory of Western Blotting

[@RohitPant04]

Quite helpful. I was confused on the part of screening but i think this video puts it together nicely. So, it's a conjugated enzyme that finally binds to the secondary antibody. On binding, the enzyme converts the colourless substrate attached to it into a colourful insoluble product that sinks down to the nitrocellulose membrane emiting colour. Got it! 💪

[@antoniomele436]

This was very helpful! Thank you!

[@CatalystUniversity]

Glad you liked it!

[@noha1878]

Great video. Thank you.

[@saharzaki2310]

But what is the point of the blocking step if the antibody would never bind to anything else?

[@叶之-p8z]

because you wouldn't see it on the membrance, colourless. And the purpose of this one experiment is to purify the protein, meaning get it out of all the crowd. When the secondary antibody binds to tthe soluble substrates and form those insoluble products, it will stick on to the memberane, and only them are the ones left.