Assay Principle
(After years of development on chemiluminescence technology, there have been a variety of luminescence systems, but the most commonly used one in laboratories is still a technology system based on Luminol or its derivatives (isoluminol, etc.). Western luminescent detection reagent ECL immunoblotting chemiluminescence solution is a non-radioactive (horseradish peroxidase) luminescence system designed to detect femtogram-level trace proteins immobilized on a solid membrane (such as NC, PVDF, etc.). It is an experimental auxiliary reagent for the photosensitive recording of its immunoblotting by X-ray film (radiograph).)
West Femto Maximum Sensitivity Substrate Reagent
West Femto Maximum Sensitivity Substrate
Applications
Western Blot, Immunoprecipitation
Synonyms
West Femto Maximum Sensitivity Substrate; N/A; Enhanced Chemiluminescence; ECL; Chemiluminescence; Electrochemiluminescence; West Femto Maximum Sensitivity Substrate reagent
Form/Format
Liquid , ready to use
Applicable Applications for West Femto Maximum Sensitivity Substrate reagent
Western Blot (WB), Immunoprecipitation (IP)
Application Notes
Highlight moment: Except the West Femto Maximum Sensitivity Substrate, there are many ways to improve the signal of immunological experiments, such as using SuperKine™ Enhanced Antibody Dilution Buffer ( ) in WB experiment, choosing DyLight, IFKine™ specialized secondary antibodies in IF experiment, or adopting high-quality animal serum, these are all pretty good options.
Usage Notes
Mix the two substrate components at a 1:1 ratio to prepare the substrate working solution.
Suggest 0.1-0.2ml of SuperLumia ECL working solution for 1 cm2 size of blot membrane.
Incubate blot 1-5 minutes in SuperLumia ECL Substrate working solution.
Drain excess reagent. Cover blot with clear plastic wrap.
Expose blot to X-ray film.
Suggest 0.1-0.2ml of SuperLumia ECL working solution for 1 cm2 size of blot membrane.
Incubate blot 1-5 minutes in SuperLumia ECL Substrate working solution.
Drain excess reagent. Cover blot with clear plastic wrap.
Expose blot to X-ray film.
Application Suggestion
1. Routine electrophoresis, membrane transfer, HRP-labeled antibody incubation, and membrane washing. It is recommended to use SuperKine™ Enhanced Antibody Dilution Buffer to dilute the antibody. Universal Loading Control Antibody Cocktail is used for the sample internal reference detection. It is recommended to use HRP-labeled IgG, such as, HRP goat anti-rabbit secondary antibody .
2. The diluted antibody should be stored at 4°C immediately after the antibody incubation, so that it can be reused later. While washing the HRP-labeled secondary antibody on the membrane, freshly prepare the luminescence working solution, and mix the two reagents at 1:1 ratio to prepare the working solution.
3. If the size of the blotting membrane is 1 cm2, it is recommended to use 0.1-0.2 ml of SuperKine™ West Femto Maximum Sensitivity Substrate working solution.
4. Incubate in the ECL working solution for 1-5 minutes.
5. Clamp the membrane with tweezers, and gently touch the lower edge of the membrane with the filter paper to remove excess luminescent liquid on the membrane. Cover the blotting membrane with a transparent plastic wrap.
6. Expose to X-ray film or take photos by chemiluminescence imager.
2. The diluted antibody should be stored at 4°C immediately after the antibody incubation, so that it can be reused later. While washing the HRP-labeled secondary antibody on the membrane, freshly prepare the luminescence working solution, and mix the two reagents at 1:1 ratio to prepare the working solution.
3. If the size of the blotting membrane is 1 cm2, it is recommended to use 0.1-0.2 ml of SuperKine™ West Femto Maximum Sensitivity Substrate working solution.
4. Incubate in the ECL working solution for 1-5 minutes.
5. Clamp the membrane with tweezers, and gently touch the lower edge of the membrane with the filter paper to remove excess luminescent liquid on the membrane. Cover the blotting membrane with a transparent plastic wrap.
6. Expose to X-ray film or take photos by chemiluminescence imager.
Precautions
1. Do not mix components from different batch numbers and different manufacturers; otherwise, it may cause abnormal results.
2. In order to obtain the best experiment results, you need to optimize all of your experiment elements, including the number of samples, antibody concentration, as well as the use of membranes and blocking reagents.
3. Mix the two substrate components at 1:1 ratio to prepare a substrate working solution. Please pay attention to change the tips during the aspiration process of A and B solution.
4. SuperKine™ West Femto Maximum Sensitivity Substrate has a long chemiluminescent duration, but it is best to perform compression or imaging within 30 minutes of color development.
5. It is recommended that every 1 cm2 membrane corresponds to 0.1-0.2 mL SuperKine™ West Femto Maximum Sensitivity Substrate working solution (femtogram level).
2. In order to obtain the best experiment results, you need to optimize all of your experiment elements, including the number of samples, antibody concentration, as well as the use of membranes and blocking reagents.
3. Mix the two substrate components at 1:1 ratio to prepare a substrate working solution. Please pay attention to change the tips during the aspiration process of A and B solution.
4. SuperKine™ West Femto Maximum Sensitivity Substrate has a long chemiluminescent duration, but it is best to perform compression or imaging within 30 minutes of color development.
5. It is recommended that every 1 cm2 membrane corresponds to 0.1-0.2 mL SuperKine™ West Femto Maximum Sensitivity Substrate working solution (femtogram level).
FAQ
1. AAA31545 can be reused? A: Repeated use is not recommended. The correct way is to add the ECL working liquid drops to the film, make it evenly covered on the film and then expose it on the machine.
2. In the process of color development, occasionally have exposure to reverse white phenomenon, what is the reason? A : This phenomenon is due to the high protein abundance, the high concentration of HRP in the central part of the chromogenic substrate will be consumed instantaneously, resulting in non-luminescence. Suggestions: (1) Reduce the loading amount of protein and the concentration of primary and secondary antibody; (2) Reduce incubation time of luminescent liquid and shorten exposure time; (3) Change to ECL color developing solution with low sensitivity.
2. In the process of color development, occasionally have exposure to reverse white phenomenon, what is the reason? A : This phenomenon is due to the high protein abundance, the high concentration of HRP in the central part of the chromogenic substrate will be consumed instantaneously, resulting in non-luminescence. Suggestions: (1) Reduce the loading amount of protein and the concentration of primary and secondary antibody; (2) Reduce incubation time of luminescent liquid and shorten exposure time; (3) Change to ECL color developing solution with low sensitivity.
Preparation and Storage
Store at 4°C, protected from light. Stable for 2 years from date of receipt.
Assay Principle
(After years of development on chemiluminescence technology, there have been a variety of luminescence systems, but the most commonly used one in laboratories is still a technology system based on Luminol or its derivatives (isoluminol, etc.). Western luminescent detection reagent ECL immunoblotting chemiluminescence solution is a non-radioactive (horseradish peroxidase) luminescence system designed to detect femtogram-level trace proteins immobilized on a solid membrane (such as NC, PVDF, etc.). It is an experimental auxiliary reagent for the photosensitive recording of its immunoblotting by X-ray film (radiograph).)
Assay Principle
(After years of development on chemiluminescence technology, there have been a variety of luminescence systems, but the most commonly used one in laboratories is still a technology system based on Luminol or its derivatives (isoluminol, etc.). Western luminescent detection reagent ECL immunoblotting chemiluminescence solution is a non-radioactive (horseradish peroxidase) luminescence system designed to detect femtogram-level trace proteins immobilized on a solid membrane (such as NC, PVDF, etc.). It is an experimental auxiliary reagent for the photosensitive recording of its immunoblotting by X-ray film (radiograph).)
Experiment Results Display
(The sample is Mouse TNF-alpha protein (, 17KD), the primary antibody is TNF-alpha Polyclonal Antibody (, 1:2500), and the secondary antibody is HRP, Goat Anti-rabbit IgG (, 1:10000). The exposure time is 30s.)
Experiment Results Display
(The sample is Mouse TNF-alpha protein (, 17KD), the primary antibody is TNF-alpha Polyclonal Antibody (, 1:2500), and the secondary antibody is HRP, Goat Anti-rabbit IgG (, 1:10000). The exposure time is 30s.)
Related Product Information for West Femto Maximum Sensitivity Substrate reagent
Principle of the Assay: After years of development on chemiluminescence technology, there have been a variety of luminescencesystems, but the most commonly used one in laboratories is still a technology system based on Luminol or its derivatives(isoluminol, etc.). Western luminescent detection reagent ECL immunoblotting chemiluminescence solution is anon-radioactive (horseradish peroxidase) luminescence system designed to detect femtogram-level trace proteins immobilizedon a solid membrane (such as NC, PVDF, etc.). It is an experimental auxiliary reagent for the photosensitive recording of itsimmunoblotting by X-ray film (radiograph).
Product Categories/Family for West Femto Maximum Sensitivity Substrate reagent
Similar Products
Product Notes
The West Femto Maximum Sensitivity Substrate (Catalog #AAA31545) is a Reagent and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's West Femto Maximum Sensitivity Substrate can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunoprecipitation (IP). Highlight moment: Except the West Femto Maximum Sensitivity Substrate, there are many ways to improve the signal of immunological experiments, such as using SuperKine™ Enhanced Antibody Dilution Buffer ( ) in WB experiment, choosing DyLight, IFKine™ specialized secondary antibodies in IF experiment, or adopting high-quality animal serum, these are all pretty good options. Researchers should empirically determine the suitability of the West Femto Maximum Sensitivity Substrate for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "West Femto Maximum Sensitivity Substrate, Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
If you are ready to order, navigate to Shopping Cart and get ready to checkout.