Can i vortex antibody

Ask Question. Asked 7 years, 8 months ago. Active 1 year, 2 months ago. Viewed times. In between each run: I switch to a new low protein binding pipette tip. I vortex the biotinylated antibody in solution in a low protein binding vial.

Then I prewet the pipette tip and deliver the reagent. Note that degradation seems unlikely. Any ideas on what may cause this? Could it be vortexing or shearing during pipette tip mixing? Improve this question. CCovey CCovey 1 1 silver badge 14 14 bronze badges. ELISA, etc.? If not, there might not be any reason to even mix the reagent. Add a comment. Active Oldest Votes. I have purchased some antibodies where the datasheet specifically says not to vortex mix. There are a few reasons why.

As a result, however, the solution takes longer to solubilize in PBS or whatever diluent you're using on its own, with no additional mechanical activity. On the other hand, even if the primary or secondary antibody is simply formulated in PBS, the equal diffusion of the proteins to all parts of the tube can take some time.

Watch it dissociate into the solution over time against a solid white background like a piece of paper. While none of these have the exact same solubility profile as antibodies, it's good enough to show you what happens over the first 30 seconds or so until you can't distinguish the dye anymore.

So, always vortex, even if it's at a relatively slow speed, for a couple of seconds to ensure everything is evenly distributed. Sign up to join this community. The best answers are voted up and rise to the top.

Stack Overflow for Teams — Collaborate and share knowledge with a private group. Create a free Team What is Teams? Learn more. Forgot to vortex antibody before staining Ask Question. Asked 5 years, 4 months ago. Active 5 years, 4 months ago. Viewed 2k times. Improve this question.

MattDMo Add a comment. Avoid bubbles and shear stress This the 1 issue we hear when people have trouble getting their protein to have function. Also, do not sonicate or pipet vigorously to a point that there are bubbles in your sample. Proteins in solution do not like air or shear stress. Add a carrier protein BSA and HSA are two common carrier proteins that increase the long-term stability of recombinant proteins. It is thought that carrier proteins help to prevent proteins from sticking to the walls of the vial or to each other.