Why tbs instead of pbs

Thanks in advance, Naomi. The whole curve OD shifted or just the high points with the 0 remaining constant? Was the M of the buffers the same? If the curve shifted did the control values remain constant?

Thanks voor your answers. But the OD of every dilution became higher. Blocking is a very important step in the immunodetection phase of Western blotting because it prevents non-specific binding of antibody to the blotting membrane. Non-fat dried milk is considered to be a good starting point when selecting a blocking solution because it is inexpensive and in very wide use.

However, milk proteins are not compatible with all detection labels, so care must be taken to choose the appropriate blocking solution for the antibodies, buffers, and detection reagents.

For example, BSA blocking solutions are preferred with biotin and AP antibody labels, and anti-phosphoprotein antibodies. This is because milk solutions contain casein, which is itself a phosphoprotein, and biotin, thus it will interfere with the assay results.

Commercially supplied blocking solutions, such as Block Ace antibody BUF from Bio-Rad, are very convenient to use and can improve consistency of results, especially when non-specific background signal is an issue.

After blocking, the blot is rinsed in wash buffer, usually TBST, with gentle agitation and in sufficient volume to keep the blot submerged. Learn more. Ask Question.

Asked 2 years, 9 months ago. Active 2 years, 9 months ago. Viewed times. Improve this question. Koreanraichu Koreanraichu 21 3 3 bronze badges. Add a comment. Active Oldest Votes. Thanks a lot in advance, I hope you can find the time to answer me.

It will be great help because I'm getting a bit out of ideas with what is going wrong with the protein I'm looking.

Thanks again. I guess that the original reason for using TBS instead of PBS is that phosphate interferes with alkaline phosphatase based detection. With HRP or fluorescent secondary ab conjugates that's not a problem. I would suggest just use PBS ph 7.