In the past, I've always used Licor's old blocking buffer that included sodium azide. I would dilute my antibodies in the buffer, stain, re-collect the dilution, and re-use it a few times.
The new Licor blocking buffer ("Intercept") says no sodium azide. Does that mean I can't re-use my antibodies? Or should I add sodium azide to the buffer?
Also, I used to always dilute the Licor buffer with PBS (1:1). Can I still do that with Intercept?
Sodium azide does not affect fluorescence. If there is no preservative, just add a little sodium azide to 0.09% final. In my experience, re-using antibody dilutions to a certain extent is fine, as long as the signal on the blot is good enough. Beware of the risk of depleting certain species in a polyclonal mix quicker than others, so your results might change over time. You might want to include a control lane in your blot as reference signal.
Regarding dilution, just give it a try and compare with undiluted. Either it works or not.
Kindly check https://www.licor.com/bio/reagents/intercept-pbs-blocking-buffer-and-diluent-kit
Hi,
I succesfully reused anti-phospho proteins 1° antibodies dilutions in LiCor Intercept Buffer. I Just recovered them after overnight incubation at 4°C, kept the in the fridge and reused them twice over 5-6 weeks (~once every 2 weeks) without sodium azide. I now favor this since a 3 times reusable 5 ml volume is much better in terms of even incubation and bubbles management than 3 times a fresh 1,667 ml. I probably could have used these dilutions 1-2 more times.
A former collaborator had a well working dilution of an antibody he kept since his post doc at least 2-3 years before, it was looking like a creepy blue-cheese solution because of aminoblack traces in partially cheesy milk. I think it was kept at 4°C and maybe they were adding some azide once in a while. Good to know it at least possible with one antibody
As Wolfgang Schechinger mentioned, azide is problematic with HRP based detection so I guess you can add some if you feel it is safer.
Best
Daniel
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