Immunofluoroscense is mainly done for detection of localization/co-localization of specific proteins, whereas Western blot is good for quantification of protein expression levels. When you talk about sensitivity, Immunofluoroscence is indeed more sensitive since it allows protein detection within a single cell whereas one can't imagine loading the protein content of one single cell for Western blot when detection level is at the microscopic level for Immunofluoroscence.

Also the antibody you use for Western blot should be able to detect a linear epitope of your protein of interest. In case you see signal with IF but get no result with WB you should check the 1 ° antibody you are using as it might not be detecting the particular conformation of your protein of interest.

In my experience, immunofluorescence yields more consistent/repeatable results. If expression levels are low, you can readily use tyramide amplification (eg, TSA from Thermo) to increase signal.

Immunofluoroscense is mainly done for detection of localization/co-localization of specific proteins, whereas Western blot is good for quantification of protein expression levels. When you talk about sensitivity, Immunofluoroscence is indeed more sensitive since it allows protein detection within a single cell whereas one can't imagine loading the protein content of one single cell for Western blot when detection level is at the microscopic level for Immunofluoroscence.

Also the antibody you use for Western blot should be able to detect a linear epitope of your protein of interest. In case you see signal with IF but get no result with WB you should check the 1 ° antibody you are using as it might not be detecting the particular conformation of your protein of interest.

If you have a lot of expressing cells, you should try western blotting.

(more than 6 cm culture dish)

If you have few cells, you should try immunofluorescence.

(1 well of 24 well plate)

file subtopics = 20 topics from the articles on " immunoflorescence staining or western blot protein expression"

may be you will find something to renew your thoughts

Each topic is represented as it's number, 20 words and 20 phrases as it is done in articles in question.

Be happy.

Western blot is more sensitive than staining. Common practice is to first see if you get it in westerns before going to tissues. There are more issues that arise in FL staining too. Background, cross talk, high antigen retrieval.

Westerns can pick up lower levels pf proteins than IHC. To make sure you have the correct bands at the proper kDa, always use a positive tissue control in your runs. Example: Pax7 = Epididymi / Caspase = stomach

I always go to protein atlas or / Labome / Antibodypedia

Thanks all for the instant help. I really appreciate it.

Immunofluorescence is generally performed for localisation and/or colocalisation of protein in cells or tissues. On the other hand, western blot (WB) is done to check the expression of a particular protein in cells or tissues. You can perform both to make your data strong. If the molecular weight of the protein is too low (below 10kDa) then you may face problems while performing WB. In that case go for IHC/IF and analyse your IHC/IF result densitometrically. If your cells/tissue of interest yields high amount of protein then WB is better. Good luck!!!