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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I also

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[@englishspoken8168]

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

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Keep uploading ..

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thanks again.. keep up..

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BaaYo Dear Madam I had applied in NIPER for VU-VIS spectroscopy, GC and HPLC chromatography Practical training short courses but there is no replay from NIPER. Can u please help me to recommend other Institutions near Chandigrah who provide me such practical training. Waiting for ur reply

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

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

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Transfer buffer
Tris base, glycine, methanol,

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