New England Biolabs Canada

Product Pathways - Companion Products

SignalFire™ ECL Reagent #6883

Item# Description List Price Web Price Qty
6883P3 SignalFire™ ECL Reagent - 50 ml $44.00
6883S SignalFire™ ECL Reagent - 500 ml $382.00
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at
Petite Kit Includes Quantity
SignalFire™ ECL Reagent A 1 x 25 ml
SignalFire™ ECL Reagent B 1 x 25 ml
Small Kit Includes Quantity
SignalFire™ ECL Reagent A 1 x 250 ml
SignalFire™ ECL Reagent B 1 x 250 ml


SignalFire™ ECL Reagent from Cell Signaling Technology (CST) is an enhanced chemiluminescent substrate capable of detecting picogram amounts of protein by western blot analysis. Compared to entry-substrates, SignalFire™ ECL Reagent boasts a more robust signal and extended duration of signal output. In the presence of hydrogen peroxide, horseradish peroxidase (HRP) converts luminol to an excited intermediate dianion. This dianion emits light on return to its ground state. Light emission is maximal immediately after exposure of the substrate to HRP and continues for 1 hour. Light can be captured on X-ray film, typically by exposure for a few seconds. Maximum sensitivity can be obtained with longer exposure.

Western Blotting

Western Blotting

SignalFire™ ECL Reagent is sensitive enough to detect picogram levels of protein. Western blot analysis of diluted recombinant human Akt-1 protein using Akt (pan) (C67E7) Rabbit mAb #4691.

Method Overview

(a) Wash membrane-bound HRP (antibody conjugate) three times for 5 minutes in TBS/T.

(b) Prepare 1x SignalFireTM Plus ECL Reagent by diluting one part 2X Reagent A (#65293) and one part 2X Reagent B (#82201) (e.g. for 10ml, add 5ml Reagent A and 5ml Reagent B). Mix well.

(c) Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film.

*Avoid repeated exposure to skin (see enclosed Material Safety Data Sheet or refer to our website for further information).


Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

Application References

  • Takebe, Y. et al. (2013) PLoS One 8, e64449. Applications: Companion Products.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


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Contents ¬©New England Biolabs Ltd.  New England Biolabs Ltd. is the exclusive Canadian distributor for Cell Signaling Technology, Inc.  New England Biolabs, Inc. is an ISO 9001 certified company.

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