Table of Contents:
1. Introduction
3. Difference Between Western Blot and ELISA
4. Is Western Blot the Same as PCR?
5. IBI Scientific: Your Western Blotting Solution
Introduction
Western blot, a key method in molecular biology and immunology, has a big impact on spotting specific proteins in complex biological samples. Scientists came up with this technique in the late 1970s. Since then, it has helped make many scientific breakthroughs and has found use in diagnosing various conditions.
Aspects of Western Blot
Sample Preparation: To get proteins from cells or tissues, you simply pull them out. Then you would scatter them up so they turn into straight lines.
Gel Electrophoresis: You split up the proteins by how big they are. You do this with a special gel that has a long name: sodium dodecyl sulfate polyacrylamide (SDS-PAGE).
Transfer: After you split them up, you move the proteins from the gel to a thin sheet. This sheet can be made of matter called nitrocellulose or PVDF.
Blocking: You don't want the antibodies to stick to the wrong places. So, you cover the sheet with a mix that has lots of protein in it. This stops the antibodies from going where they shouldn't.
Antibody Incubation: The membrane soaks in the primary antibody that targets the protein we're looking for. After washing, we add a secondary antibody. This antibody sticks to the primary one and has a special marker attached - either an enzyme or a glowing dye.
We can then spot these attached antibodies using light, glow, or color, depending on what kind of marker we used.
How We Use Western Blotting: This method has a ton of uses in research and it helps us study how proteins act, change, and work together. Western blotting also lets us check if certain proteins are in complicated mixtures.
Difference Between Western Blot and ELISA
Western blot and ELISA are both tests to find proteins, but they work in different ways and have different uses:
- How they work: Western blot sorts proteins by size using gel electrophoresis then moves them to a membrane and uses antibodies to spot them. ELISA uses a plate coated with antigens to catch proteins from a sample then uses antibodies to find them with color or light signals.
- What they're used for: Researchers often use Western blot to see if a protein is there and how much of it there is. ELISA is better for measuring how much protein is in things like blood serum because it can handle lots of samples at once on a plate.
Western Blot and ELISA are used in different ways by researchers
Is Western Blot the Same as PCR?
Nope. Western blot and PCR (Polymerase Chain Reaction) are different methods used for separate things in molecular biology:
- Western Blot: Finds and studies proteins based on their size and how specific antibodies stick to them.
- PCR: Makes more copies of specific DNA bits through steps of separating, attaching, and growing, which helps to spot and count DNA molecules.
Even though both these techniques are super important in molecular biology, they do different jobs and can't replace each other. They work together to give scientists a full picture.
IBI Scientific: Your Western Blotting Solution
Western blotting is an advanced technique that takes time, effort and patience from researchers and scientists alike. At IBI Scientific, we understand how difficult it is to find quality products for western blot at a decent price. That is why we have made it our mission to provide our western blotting tools at a 25% discount, with free shipping included.
Whether you need a standard Semi-Dry Blotter, Western Transfer Module, or a Blot Box, we have got you covered. Check out our outstanding products below:
IBI Western Blotting Products:
