WestVision[TM] Block and Diluent for Western Blots

WestVision[TM] Block and Diluent is a ready-to-use Tris buffer-based blocking solution intended for Western or dot blot applications. This solution is a proprietary formulation containing Tween 20 that improves signal intensity and resolution without increasing background. To achieve the best results, use WestVision[TM] Block and Diluent for all blocking and diluting steps including primary antibody and secondary antibody detection reagents. WestVision[TM] Block and Diluent is compatible with both alkaline phosphatase- and peroxidase- based detections systems, as well as with both chemiluminescence and chromogenic development.

To maximize the signal to noise ratio, the correct dilutions of primary antibody and secondary antibody conjugate need to be empirically determined and depend on several factors including the membrane type, the substrate to be used, the blocking/diluent solution and the amount of target. WestVision[TM] Block and Diluent is intended to be used as supplied without further dilution.

Due to the high sensitivity provided by the WestVision[TM] Block and Diluent, lower concentration of primary antibody and secondary antibody conjugates may be required to achieve optimal results.

As a starting point for chemiluminescent detection, dilute the primary antibody to 0.1 to 1.0 ug/ml and dilute the secondary antibody conjugate to 0.02 – 0.2 ug/ml, in WestVision[TM] Block and Diluent.

As a starting point for chromogenic detection, dilute the primary antibody to 0.25 to 1.0 ug/ml and dilute the secondary antibody conjugate to 0.2 – 1.0 ug/ml in WestVision[TM] Block and Diluent.

IMPORTANT: Not all blocking solutions are appropriate for all Western blot assays. Blocking solutions should be tested for compatibility in a given assay.

PROCEDURE

1. Remove the membrane from the transfer device.

2. Rinse briefly with deionized (DI) H₂O.

3. Add enough WestVision[TM] Block and Diluent to completely cover the membrane and incubate with agitation for 30 – 60 minutes.

4. Continue with your detection protocol using WestVision[TM] Block and Diluent to dilute primary antibody and detection reagents.