FAQ
What blocking agents can be used with ECL Plus Western Blotting Detection Reagents?
We recommend the ECL Blocking Agent at 5% dilution in PBST or TBST (0.1% Tween 20) buffer.
We have also found several other blocking agents which work well for ECL Plus Western Blotting on Hybond ECL and Hybond-P membranes. All produced similar sensitivities with only slightly variable signal-to-noise ratios. These chemicals (in PBST) are listed below in descending order of efficacy.
1. 5% Liquid Blocking Reagent for 30 minutes; equivalent or superior results to 5% milk.
2. 3% Bovine Serum Albumin (BSA) for 30 minutes.*
3. 1% Liquid Blocking Reagent + 0.5% BSA for 60 minutes.
4. 2% Fish Gelatin for 60 minutes.
5. 5% Normal Goat Serum for 60 minutes.
Note: BSA is the optimal choice when working with phosphorylated proteins.
How can I reprobe a chemiluminescent ECL Plus Western blot for a different target protein without stripping the blot and starting over at the blocking step? |