We often dilute antibodies in either blocking buffer or PBS (a buffer which provides physiological conditions and pH) for antibody dilution. But I am doing conjugation of commercially available antibodies with the quantum dots which I am preparing. But when I redisperse the conjugated particles in PBS after conjugation, they precipitate. My particles are stable only in water or MES buffer (its pH is 6.0). I am not sure whether my antibodies will be functional if I dilute them in water.
I don’t think antibodies would be stable in water,
I have done some particles conjugation protocols in MES buffer but always changed to PBS for storage.
Have you tried adjusting pH of your PBS? You can try acidic or basic pH (not too much) the antibodies should be stable and functional, and it can sustain particles charge and prevent precipitation.
Good luck.
Changing pH may also effect on antibody stability and binding affinity! Anyway, my suggestion is firstly you should find a suitable condition to dilute your conjugation (even in water) and then go ahead for staining with a positive control (un-conjugated antibody in same condition, as water). Make sure you get good staining in positive control -> your staining condition is ok -> if not, change other protocol like changing the pH as other people suggested above. Anyway, your positive control is important!
However, I think diluting antibody in water for just several hours staining would be ok ^^ Anyway, it needs to be answered with experiments and controls.
Good luck ^^
HI...NOT GOOD WAY .water can change the structure of antibody ..you must find the best PH for stability of yours antibody in PBS....BEST WISHES
Hi all, thank you for your suggestions. As Vu Nguyen said I have tried a control in water and it worked (luckily)... I think this antibody worked since it was kept in water only for an incubation time of 2 hours. May be not applicable for every antibodies.
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