You can do a quantitative Western, if you do it the right way. Sometimes just knowing your protein is present is all the information you need, but often you also want to know how much of your protein you have. You need to know how much protein you have For this type of information, you are going to have to run a gel and blot it. You also cannot see degradation products to determine the quality of the sample as a whole. THAT IS IT! You cannot determine the molecular weight of your protein, nor observe size shifts due to posttranslational modifications. You need more information about your protein and/or sampleĮLISAs tell you if your protein is there, and when they are done quantitatively, how much there is. Not many people have minions working for them in the lab. And if you are really lucky, you will even have an automated plate washer to help you out! Already In a microplate format, ELISAs are the assays to use for lots of samples. If you are inundated with samples, then Westerns are not for you! Western blots are more time-consuming and complex than ELISAs and unless you have a team of minions working for you, there is no way you are going to blot hundreds of samples. You have to analyze more samples than you know what to do with As long as you know the assay has enough sensitivity to detect your protein, it shouldn’t matter if you run it and blot it or drop it and adsorb it. If you just want to know if a sample contains your favorite protein, then you can use either a Western or ELISA. And what you need to know about your protein will decide which assay is best for you.īelow are some points to consider when choosing the best assay to use. So does it matter which technique you use?Įach assay has distinct advantages and differences that determine the information you can glean from your data. You stick some protein down to a support, incubate with antibodies and use a substrate that produces color, light or fluorescence to detect a protein of interest.īoth assays rely on antigen-antibody interactions to specifically detect a target protein out of a complex protein mixture. Western or ELISA – when you think about it, they are really they same assay.
0 Comments
Leave a Reply. |