I am having 2 issues with this antibodies (primary antibody from Santa Cruz Biotech):
First: The aliquots that I prepare are evaporating.
Second: They are not lasting enough to compare them with other companies.
I am aware that Santa Cruz biotech keep primary antibodies in gelatin solution instead of glycerol, so they advise not to freeze at all the antibodies. So I am trying to find out how to keep my antibodies for long periods at 4ºC or even at lower temperatures (-20 or -80ºC).
Can I make a glycerol solution to keep it from freezing or is it a bad idea?
Cheers
Ramón Lima
Gelatin is used as a protein source to protect the affinity-purified antibodies. Freezing provides aggregations between antibody and gelatin, which may alter the recognition of the antibody.
Glycerol is used to prevent antibody coupled to freeze at -20 ° c. It protects the coupling, which may break following the freezing and thawing.
To do this glycerol is used 50% final, but beware glycerol inhibits antibody if it used at a concentration of work for more than 1%.
Assuming these primary antibodies ARE NOT labeled with a reporter enzyme and you dilute them substantially (e.g. 1:100 or more) prior to use, addition of glycerol to ~10% followed by aliquoting and storage at -20 (frost free freezer) will often work with most murine subclass IgG's. IgG3s are problematic as they have a natural tendency to aggregate (multi-valent). Your mileage may vary, but some antibodies can be stored for years like this.
Thanks all for answering.
Ana, I tryed to do that but parafilm seams to dessicate after some months. I thought I could use cryotubes simmilar to those that comes with the antibodies.
Wayne I will have a closer look to those primary antibodies that I have. There are more than 15 antibodies in the same condition. Your idea is good, but my only concern is that doing exacly what you said I will dilute my antibody in the same rate I add glycerol. The other think I thought is that glycerol at 10% is not enough to keep it from freezing and the antibodiy comes with gelatin. According to Santa Cruz gelatin is the reason they do not indicate freezing they antibodies.
Lei I will make sure the temperature is sustained inside the refrigerator.
Ramón
Gelatin is used as a protein source to protect the affinity-purified antibodies. Freezing provides aggregations between antibody and gelatin, which may alter the recognition of the antibody.
Glycerol is used to prevent antibody coupled to freeze at -20 ° c. It protects the coupling, which may break following the freezing and thawing.
To do this glycerol is used 50% final, but beware glycerol inhibits antibody if it used at a concentration of work for more than 1%.
Thank you Jean-Jacques. I´ll add glycerol in our antibodies and keep it in the freezer (-20ºC). Our work solutions will be around 0,1% glycerol, so it will be safe.
Do you know why Santa Cruz choose to add gelatin in their antibodies? I don't believe this is a good choice for users.
Thanks,
Ramón
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