Antibody Dilution

[kiki06]

Hello everyone!

does anyone have a good idea how to dilute antibodies for western blot analysis?

thanks!

Kiki

[ronghai]

For me, I just dilute it in
PBS with 1%BSA, 0.05% Tween 20

[sergbio]

If your qestion about appropriate extend of dilution, so
Any antibody is unique. In many cases the activity of abs depends on their purity ( amount in mixture (polyclonal or monoclonal)) and activity ( depends on purification method and storage properties. The best choice is titration Try to start from 1:100 and till 1:10000 dilution.

[immunoassay]

For a short time, you can use PBS or PBST, too. of course, if you want to preserve Ab for a long time, you should add protectors and antiseptics into the dilution solution, including protein, polymer, glycerol, metthiolate, etc.

[admin]

For dilution factor, antibody manufacturers usually have manufacturer's instructions detailing an approximate dilution factor. Try this, then optimize.

You may be able to get more antibody usage for your money by using higher dilutions (less antibody) per western blot.

cheers

[ssacket1]

I use to dilute Ab in 0.5% skim milk in 1% TBS-T. ( 1:1000, but it depends on Ab specification)check Ab information brochure for specific Ab dilution.

[jakely]

yeah,i treat it like this.1%bovine serum,in DPBS(-)

[proteinlover]

Hi,
You can start with 1:100and go on till 1:1000.
usually I do 1:200,1:500,1:750 and 1:1000. I dilute the antibodies in 5% milk in TBS-T
Good luck!