Dear All,
Can we use Tween 20 instead of triton X in 0.1% PBT-T(I assume T stands for triton X only) for immunofluorescence ?
Also, can tween 20 be replaced by tryton in IP wash buffer?
This answer is copied from earlier discussion on researchgate under similar question that might help you
By Daniel Lee Adams If your going after a surface, extracellular, protein then there is no need to permeabilize. If the protein is intracellular or transmembrane then you need to permeabilize. Saponin, tween and triton are all good options use 0.1%-1%. Methanol or ethanol are also options if all else fails. However all proteins and cells are different, start with saponin or tween-20, then move on to the triton, then as a last resort ethanol, then methanol. This is also the order of stringency, so the saponin/tween will do the least damage to the proteins/lipids, while methanol does the most damage.
Subhash C. Juneja has posted some great info from another thread... As a rule, I usually use .1% saponin for ICC (monolayer cells on a coverslip) and .1% triton for tissue slices.
0.1% TX100 or 0.2% Tween-20 is good for immunofluorescence of most intracellular compartments. If your protein is surface expressed, no need to permeabilize. If your protein is in a dense, heavily coated intracellular compartment like a zymogen granule in an acinar cell, you may need to go up to 1-5% TX100. I know, sounds crazy.
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