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Render Timestamp: 2024-11-22T11:36:53.292Z
Commit: 5c4accf06eb7154018ba3f54329c7590f97f534a
XML generation date: 2024-09-20 06:19:53.043
Product last modified at: 2024-11-15T15:00:15.301Z
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PDP - Template Name: Detection System Kit
PDP - Template ID: *******2b9e861

SignalFire Plus ECL Reagent #12630

    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™ Plus 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™ Plus 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 room temperature. 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™ Plus ECL Reagent from Cell Signaling Technology (CST) is a highly sensitive chemiluminescent substrate capable of detecting low picogram amounts of protein by western blot analysis. SignalFire™ Plus ECL Reagent has an extended duration of signal output lasting several hours following blot exposure, allowing for multiple exposures with either film or a digital imaging system. The strong signal output allows detection of low abundance proteins, conservation of reagents, and short exposure times.
    SignalFire™ Plus ECL Reagent requires approximately five-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.
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