SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity)
(HB9308)
Technical documents: Datasheet
Product overview
| Name | SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) |
| Biological description | OverviewHello Bio SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) is a high performance enhanced chemiluminescent (ECL) substrate suitable for Western blotting of low abundance proteins in complex mixtures with horseradish peroxidase (HRP) conjugated secondary antibodies. Key FeaturesSensitivity: Equivalent to ThermoFisher SuperSignal™ West Femto Maximum Sensitivity Substrate SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) produces an excellent signal/noise ratio with low background to produce publication quality Western blots. NotesWe recommend:
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| Customer comments | Works beautifully, as always with any Hello Bio product! We tested this ECL Western Blotting substrate kit and it works marvelously. We get comparable result to our previous supplier's kit and are committed to switching to Hello Bio products for our ECL WB. Thank you Hello Bio as always for a wonderful product! Verified customer, Aston University I used your ECL solution for a trial and it worked well on my blots, I liked it! Less background and less incubation time which was great. Verified customer, University of Bristol |
| Description | High sensitivity ECL solution for developing chemiluminescent Western blots |
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Images
Figure 1. Representative blots for Neurofilament L of SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) and competitor solutions
Representative blots of Hello Bio ECL solutions and competitors showing SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) providing extremely high sensitivity with low background. Method: A 12% acrylamide gel was loaded with a dilution series of rat brain cytosol fraction (10µg to 5ng total protein per lane) and transferred to a PVDF membrane. This was then probed with a monoclonal rabbit anti-neurofilament L antibody (HB7266, 1:10,000 dilution, 100ng/ml) primary antibody and goat anti-rabbit HRP (1:10,000 dilution) secondary antibody. The blot was exposed with every ECL solution (500µl part A + 500µl part B) using a Licor Odyssey Fc digital imager . For more information on Western Blotting please see our Western blotting protocol.
Figure 2. Fully counterbalanced comparison of Hello Bio ECL substrates with competitor products.
SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) shows extremely high sensitivity equivalent to Thermofisher SuperSignalTM West Femto Maximum Sensitivty Substrate. Method: 12% acrylamide gels were loaded with a dilution series of rat brain cytosol fraction (10µg to 5ng total protein per lane) and transferred to PVDF membranes. These were then probed with a monoclonal rabbit anti-neurofilament L antibody (HB7266, 1:10,000 dilution, 100ng/ml) primary antibody and goat anti-rabbit HRP (1:10,000 dilution) secondary antibody. All blots were exposed with every ECL solution (500µl part A + 500µl part B) in a fully randomised Latin square design to avoid any order effects. ECL exposure was carried out following the protocol detailed in "Application Notes". Imaging was carried out using a Licor Odyssey Fc digital imager and all band intensities were calculated in Licor Image Studio. For more information on Western Blotting please see our Western blotting protocol.
Figure 3. Dot blot for neurofilament L in a complex protein mixture (rat brain cytosol fraction) using SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity)
SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) is able to detect Neurofilament L in as little as 1.5pg of rat brain cytosol fraction. Method: A nitrocellulose membrane was spotted with a dilution series of rat brain cytosol fraction (450ng to 1.5pg). Following blocking in 5% non-fat dry milk the membrane was probed with a monoclonal rabbit anti-neurofilament L antibody (HB7266, 1:10,000 dilution, 100ng/ml) primary antibody and goat anti-rabbit HRP (1:10,000 dilution) secondary antibody. The blot was exposed with SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) (750µl part A + 750µl part B) using a Licor Odyssey Fc digital imager.
Figure 1. Representative blots for Neurofilament L of SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) and competitor solutions
Representative blots of Hello Bio ECL solutions and competitors showing SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) providing extremely high sensitivity with low background. Method: A 12% acrylamide gel was loaded with a dilution series of rat brain cytosol fraction (10µg to 5ng total protein per lane) and transferred to a PVDF membrane. This was then probed with a monoclonal rabbit anti-neurofilament L antibody (HB7266, 1:10,000 dilution, 100ng/ml) primary antibody and goat anti-rabbit HRP (1:10,000 dilution) secondary antibody. The blot was exposed with every ECL solution (500µl part A + 500µl part B) using a Licor Odyssey Fc digital imager . For more information on Western Blotting please see our Western blotting protocol.
Figure 2. Fully counterbalanced comparison of Hello Bio ECL substrates with competitor products.
SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) shows extremely high sensitivity equivalent to Thermofisher SuperSignalTM West Femto Maximum Sensitivty Substrate. Method: 12% acrylamide gels were loaded with a dilution series of rat brain cytosol fraction (10µg to 5ng total protein per lane) and transferred to PVDF membranes. These were then probed with a monoclonal rabbit anti-neurofilament L antibody (HB7266, 1:10,000 dilution, 100ng/ml) primary antibody and goat anti-rabbit HRP (1:10,000 dilution) secondary antibody. All blots were exposed with every ECL solution (500µl part A + 500µl part B) in a fully randomised Latin square design to avoid any order effects. ECL exposure was carried out following the protocol detailed in "Application Notes". Imaging was carried out using a Licor Odyssey Fc digital imager and all band intensities were calculated in Licor Image Studio. For more information on Western Blotting please see our Western blotting protocol.
Figure 3. Dot blot for neurofilament L in a complex protein mixture (rat brain cytosol fraction) using SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity)
SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) is able to detect Neurofilament L in as little as 1.5pg of rat brain cytosol fraction. Method: A nitrocellulose membrane was spotted with a dilution series of rat brain cytosol fraction (450ng to 1.5pg). Following blocking in 5% non-fat dry milk the membrane was probed with a monoclonal rabbit anti-neurofilament L antibody (HB7266, 1:10,000 dilution, 100ng/ml) primary antibody and goat anti-rabbit HRP (1:10,000 dilution) secondary antibody. The blot was exposed with SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) (750µl part A + 750µl part B) using a Licor Odyssey Fc digital imager.
Biological Data
| Application notes |
Protocol for Chemiluminescent blot development with ECLDigital Imaging
Film Imaging
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Solubility & Handling
| Storage instructions | +4°C (protect from light) |
| Storage buffer | Contains 0.05% ProClin-300 |
| Important | This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use. |
Publications
These publications cite the use of SuperBlotTM ECL Western Blotting Substrate Kit (High sensitivity) purchased from Hello Bio:
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Novel microneedle patches for transdermal delivery of AP39, a hydrogen sulphide donor, in the treatment of scenarios mimicking neurological disorders
Balakrishnan et al (2024) Journal of Pharmaceutical Investigation : https://doi.org/10.1007/s40005-024-00711-9
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