Protocol Online logo
Top : Forum Archives: : Protein and Proteomics

Reuse antibodies for further incubation in Western blot - (Mar/16/2005 )

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

-mac1-

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

Simon

-Simonsays-

QUOTE (Simonsays @ Mar 16 2005, 01:59 PM)
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 tongue.gif

-mabusheh-

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.

-fred_33-

QUOTE (fred_33 @ Mar 17 2005, 03:13 AM)
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 cool.gif
use it at 0.02% it works fine

-mabusheh-

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.

-Nay-