Both detergents are structurally very similar, but Triton X114 has a shorter polyoxyethylene headgroup than X100 (7-8 vs 9-10 units). As a consequence it has a higher cmc (350 vs 50 µM) and a lower cloud-point (22 vs 65 °C). Both are viscous fluids, their handling is quite similar. Both can form peroxides, so they need to be stored tightly capped, cool and protected from light. Removal of peroxide is possible, but tedious and requires problematic solvents. It is generally better to buy peroxide-free rather than the (cheaper) industrial preparations.

More specific answers would require knowledge of your protocol.

Thanks Dr Buxbaum. I also found that though both can form homogenous phase at lower temperature with water,Triton X-114 will form a biphase at a higher temperature.That is why it prefered in lysis buffer since the desired component can disole into one of the phases and can easily be isolated.

Is there any method that can make Triton X-100 form a biphase so that aqueous phase can be distinct from the detergent phase?

Yes, you need to heat the mixture above the cloud point. As I pointed out, that is much higher in X100 than in X114, which is why x114 is used for this application.

Thank you Very much for your erudite answer.First of all I needed to prepare 32% Triton X-114 but since I Have Triton X-100 I want to make use of it to prepare my breaking buffer.What Protocol do you suggest tme.Thanks again.

Because Triton detergents are so viscous, I weigh the detergent (that is, prepare solutions by weight% rather than volume%). Once you mixed it thoroughly with some water, viscosity drops and you can add the other buffer components and finally make up to the desired volume. Recall though that polyoxyethylene detergents are subject to oxydation, so exclude air as much as possible.

Thanks again.I had a mono phase mixture of Triton X-100 and water at room temperature but when I heated it at above 60 degrees Celsius I got a biphase.

Yes, of course. The cloud point of triton x100 is 63 °C, above that temperature it no longer mixes with water. That is the whole point of using x114, the cloud point is lower (25 °C), so hydrophobic proteins can be extracted into the detergent phase (Article Triton X-114 based cloud point extraction: A thermoreversibl...

Engelbert Buxbaum. Sir, Can I use TX114 for total protein extraction from bacterial cell? If yes, How much efficient it is in comparison to other methods including urea and SDS coupled buffers? How much concentration need to use?

Polyoxyethylene detergents like the triton are non-ionic and relatively mild. For example, detergent-resistant membranes (DRM) is a triton-insoluble fraction of the plasma membrane, overlapping with rafts (but not completely identical). Because they are uncharged, they are useful for IEF (in combination with (thio-)urea), but for conventional electrophoresis I'd use either SDS or, especially if I am interested in glycoproteins, CTAB (doi:10.1016/S0003-2697(02)00639-5). If you are interested in detergents and membrane proteins, I have reviewed them in ISBN 9781441972507.