Femto 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 stable for 12 months.    These products are shipped at RT.

Introduction

  The Fluorescence West Femto ECL Substrate is an extremely sensitive enhanced chemiluminescent substrate for detecting horseradish peroxidase (HRP) on immunoblots. This substrate’s extremely intense signal output enables detection of low femtogram amounts of antigen. Furthermore, this substrate requires substantial dilutions of the primary and secondary antibodies. The sensitivity, intensity and duration of the signal allow for easy detection of HRP using photographic or other imaging methods. Blots also can be repeatedly exposed to film to obtain optimal results or stripped of the immunodetection reagents and reprobed.

Procedure Summary

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

1.           Dilute primary antibody to 5-100ng/mL.

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

3.           Mix the two-substrate components at a 1:1 ratio to prepare the substrate Working Solution.

4.           Incubate blot 5 minutes in West Femto 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.