Blot data in 1ST in a few clicks

Aug 6th, 2022
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How to blot data in 1ST

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this video is Western blot I want to make a couple points about why we are studying this first off this is a key technique that we use to visualize proteins so this is one of our goals by learning about this technique weamp;#39;re going to reinforce some of our knowledge about protein chemistry secondly weamp;#39;re going to learn how to interpret data from a Western blot which is a course theme and weamp;#39;re gonna go through the first few learning outcomes in this video so a Western blot is a method to detect specific proteins within a sample or tissue it can tell us about the presence and the amount of the protein this is a Western blot here so it looks a lot like a DNA gel and this is somewhat analogous to agarose gel electrophoresis as a way to visualize DNA however in this technique weamp;#39;re using a Western blot which will break into several techniques to visualize protein now I want to show you how to break down a Western blot so that we can get specific results about

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A western blot is a laboratory method used to detect specific protein molecules from among a mixture of proteins. This mixture can include all of the proteins associated with a particular tissue or cell type.
To know how to analyze western blot data, Look for the sizes of the bands. These will be represented by a number, either followed by kDa or preceded by p. This is the size of the protein which has been detected and is the scale on which the proteins are separated in a Western blot.
To perform a Western Blot successfully, every single step should not be neglected. It includes: (1) WB buffers preparation, (2) samples preparation, (3) gel electrophoresis, (4) protein transfer, (5) membrane blocking, (6) antibody incubation, (7) WB detection and imaging, (8) Data analysis.
Western Blot Presentations Label every lane (optional: use numbers and have a number key below) Label ladder band sizes. Label band(s) of interest with protein name. Label control band with protein name.
To this end, once a western blot has been imaged, it can then be analyzed. To do this, researchers use a technique called densitometry to measure the relative amount of a specific protein for a given experimental sample on the blot by comparing it with a control or another protein.
Western Blotting (WB) The membrane is then blocked for 1 hour. Afterwards, it is incubated with primary antibody for 2 to 12 hours overnight followed by the secondary antibody incubation, lasting another hour.
A Western blot test is typically used to confirm a positive HIV diagnosis. During the test, a small sample of blood is taken and it is used to detect HIV antibodies, not the HIV virus itself.

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