Composition of Antibody dilution buffer

[jagan]

Dear all,

The Antibody dilution buffer presently what i am using is 1% BSA, 0.1% Cold Fish Skin Gelatin, 0.05% Sodium Azide in 10mM PBS. But my Antibody is not stable in this buffer, If i am freshly preparing means it is giving the Result in IHC. When i kept for stability for 1weak its not giving the result. So i want the different Composition of Antibody dilution buffer. Can u suggest in this aspect?

Thanks and Regards,

Jagan

[CanuckPhD]

My dilution buffer for the primary antibody is 1% of the blocking serum in Tris (50mM) buffered saline (150mM). Example if I block in 10% goat serum, then the antibody dilution buffer is 1%  goat serum. It is made up fresh every time I use it and the primary added at that time.

I have found that it gives me as good results as when I used to use a more complex buffer of BSA, serum and casein.

Remember that diluted antibody solutions often do not last if stored. For free floating I have been able to use diluted solutions if I froze them for less then 1 month.

Good luck

[jagan]

Dear Canuck

The Antibody dilution buffer what i am using now is 1% BSA, 0.1% Cold Fish Skin Gelatin, 0.05% Sodium Azide in 10mM PBS i am satisfied with this when i am using freshly, but my problem is Stability. I want the Antibody dilution buffer composition which is stable for long time.

Thank you for your valuable suggestion,

Regards

Jagan

[CanuckPhD]

If you use 1% blocking serum as your antibody dilution buffer it is easy and quick. I make it up fresh each time and it works well. It only takes about 1 minute to make a 1:10 dilution of the 10% serum blocking buffer.

This way you do not need to have a stable dilution buffer.