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Amersham ECL Plus™ Western Blotting Detection Reagents- High sensitivity: broad dynamic range on film and imager, which allows a quantitative detection.
- Chemiluminescent and chemifluorescent detection.
- Extended signal duration¾up to 24 h¾enables multiple exposures to be made.
- Compatibility with Hybond™ ECL™ and Hybond-P membranes.
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Amersham ECL Plus™ Western Blotting Detection Reagents
Technical Information
Amersham™ ECL Plus Western Blotting Detection Reagents deliver superior sensitivity and a broad dynamic range. The acridan-based substrate used in ECL Plus Western Blotting Detection Reagents releases a high level sustained output of light. In addition, ECL Plus Western Blotting Detection Reagents generate an intense long-lasting chemifluorescent signal at 440 nm that can be detected on a fluorescence imager with the appropriate filters.
This versatile functionality enables you to carry out qualitative analysis (on film) and quantitative analysis using CCD imagers or laser scanners.
RPN2132 contains the following reagents, sufficient for detecting 1000 cm2 membrane:
- ECL Plus Solution A, 100 ml
- ECL Plus Solution B, 2.5 ml
RPN2133 contains the following reagents, sufficient for detecting 3000 cm2 membrane:
- ECL Plus Solution A, 300 ml
- ECL Plus Solution B, 7.5 ml
A
B
Target: mouse IgG slot blotted onto PVDF (Hybond-P, RPN 2020F); loadings: doubling dilutions starting at 5ng; detection: 1:2500 dilution of anti-mouse Ig HRP conjugate (NA 931) using (a) ECL detection reagents (RPN 2106) and (b) ECL Plus detection reagents (RPN 2132); exposure : on Hyperfilm™ ECL (RPN 2103) for 5 min.
Target: rat brain homogenate Western blotted onto PVDF (Hybond-P, RPN 2020F); loadings: doubling dilutions starting at 1:20 dilution of the homogenate; detection rimary antibody - 1:5000 dilution of mouse anti-beta tubulin (N 357), secondary antibody - 1:5000 dilution of anti-mouse Ig HRP conjugate (NA 931) using ECL Plus detection reagents (RPN 2132); exposure: on Hyperfilm ECL (RPN 2103) for 60 min, 24 h after initiation of the detection reaction.
25 50 75 100 125 150 µg
A
B
Western blot analysis of bcl-2 using ECL, and ECL Plus. Gels were loaded with between 25 to l50 µg of total heart protein. Both blots were exposed to Hyperfilm ECL for 15 min and film result scanned by document scanner; (a) ECL standard reagents, (b) ECL Plus reagents. Courtesy of Dr M J Dunn and Dr N Latif, National Heart & Lung Institute, Imperial College School of Medicine, Harefield Hospital, Middlesex.
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Product related documents
Application note
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; loadings: doubling dilutions starting at 1:20 dilution of the homogenate; detection rimary antibody - 1:5000 dilution of mouse anti-beta tubulin (N 357), secondary antibody - 1:5000 dilution of anti-mouse Ig HRP conjugate (NA 931) using ECL Plus detection reagents (RPN 2132); exposure: on Hyperfilm ECL (RPN 2103) for 60 min, 24 h after initiation of the detection reaction.'))
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