5 replies to this topic

[mac1]

I have a question on the usage of antibodies in general for western blotting. How many times can one reuse the primary and secondary antibodies?

Please share from your experiences.

Many thanks

Edited by bioforum, 22 March 2005 - 11:28 PM.

[Simonsays]

I reuse them as long as the signal stays good. You can't reuse is though if you incubate with BSA or milk.

Simon

[mabusheh]

Simonsays, on Mar 16 2005, 01:59 PM, said:

I reuse them as long as the signal stays good. You can't reuse is though if you incubate with BSA or milk.

Simon

I reuse it as long as the signal stays good even when I incubate with 1% BSA or milk. I keep it at 4 degrees fridge with 0.02% Na Azide.
and it works fine, real fine

[fred_33]

hi
i reuse antibodies until the signal becomes reared. But if the milk stinks, i waste the solution.

Quote

0.02% Na Azide

is that product to prevent the milk degradation?
i use to store my antibodies solutions at -20°....but if i add Na azide should i store at 4 or -20 is still fine?

Thanks in advance.

Edited by fred_33, 17 March 2005 - 02:15 AM.

[mabusheh]

fred_33, on Mar 17 2005, 03:13 AM, said:

hi
i reuse antibodies until the signal becomes reared. But if the milk stinks, i waste the solution.

Quote

0.02% Na Azide

is that product to prevent the milk degradation?
i use to store my antibodies solutions at -20°....but if i add Na azide should i store at 4 or -20 is still fine?

Thanks in advance.

Hi
Na Azide prevents bacterial growth, so it won't stink for longer time
use it at 0.02% it works fine

[Nay]

I store my Ab's diluted in milk - I find it is better to dissolve the milk powder in autoclaved/sterilised PBS before adding the antibody too - this helps it keep for longer at 4 degrees.