Antibody Technical Tips

 

Tip 1:  Use of antibodies in western blotting

Tip 2: Western blotting vs immunochemistry

 

For more specific technical tips please view our product pages and datasheets. 


 

Tip 1: Use of antibodies in western blotting

Tech Tip #1:  To improve detection of the protein of interest, dry the PVDF membrane after transfer for 15-20 min.  This improves binding of the protein onto the PVDF membrane.  This has not been tested with nitrocellulose membranes.  After drying, the PVDF membrane is re-activated with 100% methanol and incubated with blocking solution.

Tech Tip #2:  When possible, include a positive control sample on the western blot to confirm that the antibody is able to detect the protein of interest.

 

Tip 2: Western blotting vs immunochemistry

The same antibody does not always work well for both western blotting and immunochemistry.  And even when the same antibody is compatible with both techniques, often different antibody dilutions need to be used.  Determining the optimal antibody dilutions needs to be done empirically or based on reports in the literature with the same antibody.