Western blotting technique | Principle | Step by Step | In Hindi

This lecture explains about the western blotting technique step by step in Hindi.
The Western blot is meant for detecting proteins. The proteins are first purified from cells and then separated. During the separation you need to choose whether to keep the proteins natural or denature them. Denatured proteins are often easier to separate, but you might be looking for morphological variations. Separation is usually done on a gel followed by electrophoresis.
Often the gel will be stained after the electrophoresis so the proeins are visible. This is done to make sure that the electrophoresis worked.
Then the proteins are blotted from the gel and over to a membrane, usually nitrocellulose, Immediately after the blotting the membrane are blocked so no other proteins will attach themselves to the membrane.
After the blotting the membrane is induced so the proteins can be detected. Usually specific antibodies for the exact protein you are working with will be used.
A secondary antibody that can bind to the first antibody is then added and the secondary antibody has some kind of detection mechanism, either fluorescence, enzyme or color.
Please watch this video to have a clear idea about 2D gel electrophoresis.
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[@manojsharma-vn4ij]

Simple and classy way of explaining..... really you explained not just protocol but the concept behind it..thanks for the efforts......best wishes for upcoming videos.

[@KARZDAAR_MUSIC]

Tomorrow is my presentation on western blotting technique in college, hope this helps :( I'm memorizing this 🙂

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How can you explain such a complex method into such simple language ❤

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[@BaaYo]

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[@akhileshkumarmaurya1240]

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I also

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Good afternoon ma'am. Welcome again after long time.

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On of my best channel simply explanation from Pakistan 😊

[@englishspoken8168]

1. Isolate protine 2 sample preparation 3. Sds page 4. Transfer on blotting pape Nitrocellulose and PVDF membrane

[@JYOtiRaNJanMANgaRaj]

THANK U VEY MUCH GOOD UNDERSTANDING 🙏👌👌🙂🙂

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This video is really amazing and helped me to understand the western blot. Thank you very much 😊

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[@sureshgoswami177]

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Very good ma'am, thank you so much Ma'am.

[@dikshantkumar7660]

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[@aniketsarkardava]

Madam,what kinds of protein detection method can be used in western blotting ?

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I am in class 12 this helps me a lot .....👍👍

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[@muzamilarshad2671]

i have a question. why proteins are transferred from gel to membrane? function of secondary antibody?

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As always very much helpful mam

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Mam, I want to know, is it require to again activate the PVDF membrane after the transfer?

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[@BaaYo]

Sure..will try to upload before 16th june..

[@paramitachakravarty2714]

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Why don't we make primary antibody labelled ?

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Mam inke naam Western, southern, northern blotting he kyu h

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Transfer buffer Tris base, glycine, methanol,

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Mam I hv lil bit queries like Why we don't use Enzymes tagged primary anybody instead used of secondary anti primary Ab!!??

[@suganyasivakumar6972]

It’s for the ease of use .. as everytime we develop primary antibody against the protein of interest in mouse preferably.. we don’t have to tag it with enzyme .. and the secondary antibody is going to bind to the constant region of the antibody .. so mostly .. the fab region of the antibody only is going change .. and the constant region doesn’t vary between different clones of antibody .. so the secondary tagged antibody can be purchased which is commercially available .

[@Entertainmentplus67]

@@suganyasivakumar6972yes, that I can understand. But my question was simple, can we use direct enzyme tagged primary Ab (if it commercially available/some company providing it by request!!)

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👌

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Bovine serum albumin :)

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Mam please make a video on eastern blotting

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platelet aggregation test maam ,can u plz share