Dear Tareg,

In my opinion, PBS with 1% BSA as washing buffer contains too much BSA (I use such concentration for blotting techniques). In my experience with bead-based immunoassays PBS with 0.1% BSA was sufficient. I would recommend you to compare results of some commonly used test/experiment with commercial buffer then PBS with 1% BSA buffer and PBS with 0.1% BSA buffer. You will see if some differences occure. If it does not, 0.1% BSA in PBS is sufficient. If it does even in 1% BSA, some other supplements might be considered (e.g. tween 20).

Good luck

Zuzana

Dear Tareg,

we apply the bead-based Luminex technology for about two years. In the first time we varied the composition of the washing and dilution fluids:

1. As dilution fluid we use PBS, 1% BSA, 0,02% Tween 20, 0,1% NaN3 (for dilution of sample, anti Ig, Streptavidin-Phycoerythrin)

2. As washing fluid for the microtiter plates we also use PBS, 0,02% Tween 20, 0,1% NaN3

3. As washing fluid for the Luminex200 instrument we always use “Flow Cytometry, BD FACSFlow Sheath Fluid”

Pos. 1 and 2 were filtered by pore size of 0,2µm.

I hope, these details will help you in your research.

With best regards

Joachim Hoernig

Tareg, 

we nominally buy buffer separately about 1L and  add about 20-25% of DI water before each use.

Thank you all for the insigftul suggestions. Today I just tried the PBS, 1% BSA buffer and although the generated FI was not adversely affected, there was much bubble produced. Next time I will try to see what good use I may be able to make of your valuable suggestions. I would be happy if Nikunj Gevariya can take the time to let me know the details of the buffer and some recommended online vendor.

Hi Tareg,

We buy Bio-Plex® Wash Buffer #171-304500.