Pico ECL^® Western Blotting Substrate

DuraECL^®, FemtoECL^®，PicoECL^®，ECLplus^®are registered trademarks and strictly forbidden to be used without authorization.

Kit Contents:  ECL Solution A, 10mL; ECL Solution B, 10mL;  sufficient for 200cm²of membrane

Storage: Upon receipt store solutions at 4°C for 12 months.  These products are shipped at ambient temperature.

Introduction

    The Pico ECL Substrate is a highly sensitive nonradioactive, enhanced luminol-based chemiluminescent substrate for the detection of horseradish peroxidase (HRP) on immunoblots. The Pico ECL Substrate enables the detection of picogram amounts of antigen and allows for easy detection of HRP using photographic or other imaging methods. Blots can be repeatedly exposed to X-ray film to obtain optimal results or stripped of the immunodetection reagents and re-probed. The optimally formulated Pico ECL Substrate makes it the ideal substitute for other similar substrates without the need for additional optimization of assay conditions.

Procedure Summary

      Note: The recommended range of antibody dilutions should be used as references to obtain positive results. The optimal antigen and antibody amounts to be used may require experimentation.

1.           Dilute primary antibody to 100 - 500 ng/mL.

2.           Dilute secondary antibody to 10 - 50 ng/mL.

3.           Mix the two substrate components (A and B) at a 1:1 ratio to prepare the substrate Working Solution.

4.           Incubate blot 5 minutes in West Pico ECL Substrate Working Solution.

5.           Drain excess reagent. Cover blot with clear plastic wrap.

6.           Expose blot to X-ray film or Use imaging devices.

NOTE: Always wear lab coats, gloves and goggles when working with our products although they are low-risk chemicals for R&D only.

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