Render Target: STATIC
Render Timestamp: 2024-11-21T12:53:08.141Z
Commit: 5c4accf06eb7154018ba3f54329c7590f97f534a
XML generation date: 2024-09-20 06:19:56.450
Product last modified at: 2024-11-15T21:30:11.314Z
1% for the planet logo
PDP - Template Name: Detection System Kit
PDP - Template ID: *******2b9e861

SignalFire Elite ECL Reagent #12757

    Product Information

    Product Usage Information

    (a) Wash membrane-bound HRP (antibody conjugate) at least three times for 5 minutes in TBS/T. It is very important to thoroughly wash the membrane prior to substrate incubation.

    (b) Prepare 1X SignalFire™ Elite ECL Reagent by diluting one part 2X Reagent A and one part 2X Reagent B (e.g. for 10 ml, add 5 ml Reagent A and 5 ml 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).

    Solutions and Reagents

    Each SignalFire™ Elite ECL Reagent (A and B) is a 2X concentrate; there is no need to further dilute in water when the two reagents are combined.

    Storage

    Store at 4ºC. This product is stable for 12 months. DO NOT store Reagents A and B pre-mixed. Reagents A and B should be combined just prior to exposing membranes.

    Product Description

    SignalFire™ Elite ECL Reagent from Cell Signaling Technology (CST) is an ultra sensitive chemiluminescent substrate capable of detecting femtogram amounts of protein by western blot analysis. SignalFire™ Elite ECL Reagent is compatible with both film and digital imaging systems. The extremely intense signal output allows detection of very low abundance proteins, conservation of reagents, and short exposure times.
    SignalFire™ Elite ECL Reagent requires approximately ten-fold less Anti-rabbit IgG, HRP-linked Antibody #7074 or Anti-mouse IgG, HRP-linked Antibody #7076 than traditional ECL reagents. Limiting the amount of HRP exposed to the membrane prevents high background, oversaturation of the target protein signal, or false negative results. Other HRP-conjugated antibodies, including HRP-conjugated primary and anti-biotin-HRP antibodies, should be diluted similarly. Dilution of secondary antibody from alternative vendors may need to be optimized. Titration of lysate and primary antibody concentration is recommended to achieve optimal signal-to-noise ratio.

    Background

    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.
      For Research Use Only. Not For Use In Diagnostic Procedures.
      Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
      All other trademarks are the property of their respective owners. Visit our Trademark Information page.