eBioscience
Intracellular Fixation & Permeabilization Buffer Set
88-8824-00
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The specific aim for using this product was to permeabilize T cell membranes to allow intracellular detection of FoxP3 using fluorescent antibodies. I chose this product because it both fixes and permeabilizes at the same time.
Membrane permeabilization for Flow Cytometry
Regulator T cells isolated from peripheral blood
I isolated T cells from peripheral blood of control and treated mice to look at changes in T cell markers associated with drug treatment. I then stained with various extracellular cell surface marker antibodies, followed by permeabilization using this kit to allow intracellular staining of FoxP3.
The protocol suggests that you can go for as little as 30 minutes, but I have found that I need to go at least an hour to get successful permeabilization of the membrane for intracellular staining.
I was able to successfully stain for intracellular markers using this perm/fix kit. It was easy to use and the stock solutions are easy to prepare.
Sometimes the buffers will get some precipitate in the bottle, so you will need to warm the solution back up (37 degrees) to dissolve it again.
Easy to use, inexpensive, lasts a long time
Takes longer than the protocol states
The kit is easy to use, and I would recommend it for membrane permeability followed by flow cytometric analysis!
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