Who have used beads covered with protein in ELISA? - Are they heavy enought to be washed out? (Dec/14/2005 )
I have some problem with my 0.1 um beads. I have conjugated protein to it, and wanted to check out with ELISA, if this protein is reactive , on the beads. And I had no signal. In the control wells, with the protein alone, signal was pretty fine.
I just wonder, coud the beads be too heavy to be washed out during washing steps? But they are very very small, 8 times more small than the bacteria E.coli, for example.
-Alesia-
QUOTE (Alesia @ Dec 14 2005, 10:52 AM)
I have some problem with my 0.1 um beads. I have conjugated protein to it, and wanted to check out with ELISA, if this protein is reactive , on the beads. And I had no signal. In the control wells, with the protein alone, signal was pretty fine.
I just wonder, coud the beads be too heavy to be washed out during washing steps? But they are very very small, 8 times more small than the bacteria E.coli, for example.
I just wonder, coud the beads be too heavy to be washed out during washing steps? But they are very very small, 8 times more small than the bacteria E.coli, for example.
Hi Alesia,
Seems like you are having quite a few problems in lab today!!
Are your beads magnetic? If so, use the magnet to hold them while you wash. Also, with some beads, they can be spun in microcentrifuge tube and the wash pipetted out away from them.
If you are using them in an elisa tray, you will risk pulling them up with the wash buffer.
Good Luck,
Cindy
-biokmst-
Thank you for answer.
No, my beads are not magnetic.
Is it possible than to do something like ELISA sandvich, but not on the plate, in tube, in centrifuging every time ? It could take quite a time, because to pellet so little beads I need to centriguge them for 1 h. But I will try maybe, since ELISA does not work...
-Alesia-