Blot data in NB in a few clicks

Aug 6th, 2022
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How to blot data in NB

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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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Western Blot Western blotting is a laboratory technique used to detect a specific protein in a blood or tissue sample. The method involves using gel electrophoresis to separate the samples proteins.
Western blot is a reliable quantitative method only if sample properties and integrity, antibody specificity to the target protein, and loading protocols are considered. With careful attention to details, you can avoid common mistakes and avoid misinterpreting Western blot data.
During the test, a small sample of blood is taken and it is used to detect HIV antibodies, not the HIV virus itself. The Western blot test separates the blood proteins and detects the specific proteins (called HIV antibodies) that indicate an HIV infection.
A qualitative Western blot shows the presence or absence of a protein of interest but provides no information on relative protein abundance. A quantitative Western blot is used to detect specific proteins and measure relative protein abundance.
It detects viral antigens (proteins usually on the surface of viruses) using antibodies against those proteins. A positive Western blot indicates the presence of viral antigen which very often means live virus in our patient. That patient may have an ongoing viral infection.
Quantification. It is very important to be aware that the data produced with a western blot is typically considered to be semi-quantitative. This is because it provides a relative comparison of protein levels, but not an absolute measure of quantity.
Despite this proliferation of blotting names, other than the Southern blot, none of them is actually named after a person. Therefore, except for Southern blot, the word that precedes blot should not be capitalized.
Protocol for Quantitative Western Blots Choose a Normalization Strategy. Prepare Samples. Perform Electrophoresis. Transfer to Membrane. Block. Incubate with Primary Antibodies. Wash. Incubate with Near-Infrared Fluorescent Secondary Antibodies.

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