Diluting antibody stocks in glycerol
Posted 21 March 2012 - 10:58 AM
Posted 21 March 2012 - 11:51 AM
Posted 22 March 2012 - 01:31 AM
Posted 22 March 2012 - 11:35 AM
Diffusion rate is definitely a phenomenon...
Posted 22 March 2012 - 02:30 PM
I will try to make sure to make a correct dilution (1:1 glycerol-antibody). Ultimately, the staining is what I'm worried about since glycerol could interfere with the visualisation (or not)?
And yes, the main reason for doing this is so that antibody stocks doesn't freeze at -20 C since we will be freeze-thawing it regularly. I've also aliquoted them into aliquots.
Posted 23 March 2012 - 12:01 AM
Posted 23 March 2012 - 12:57 AM
"Some researchers add the cryoprotectant glycerol to a final concentration of 50% to prevent freeze/thaw damage; glycerol will lower the freezing point to below -20oC. While this may be acceptable for many antibodies, only a small percentage of the antibodies we offer have been tested for stability in this storage condition and our guarantee only applies to antibodies stored as recommended on the data sheet."
Which would make me think that it would might be good idea to check if using glycerol will have an effect on your staining empirically. I don't see why they would make such a claim if it hasn't been a problem for someone along the way.
Although as doxorubicin states, many antibodies do come in buffers containing glycerol, so the likelihood of any adverse effect is probably low.
(bob1, I too read and understood the original post as you did..... )
Posted 23 March 2012 - 09:55 AM
you have to remember, though, that the antibody concentration will now be half that of the original. instead of using a 1:100 dilution you will now need to use a 1:50 dilution of the glycerol stock to get the same concentration of antibody in your working solution.
genius does what it must
i do what i get paid to do