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Durastab Medtec LTD has always adhered to the mission of "making diagnostic reagents more stable and more accurate" since its establishment, and is committed to providing excellent ready-to-use reagent solutions to global advanced medical diagnostic companies.
Until now, Durastab has provided products or services to more than 600 companies and 500 scientific research institutes, of which more than 50 IVD companies are listed companies in China or overseas.
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Flagship Product: Safer, More Stable, and More Versatile – ProCide™ 300 Preservative!
Newly Upgraded 3rd-Gen TMB Substrate: Enhanced Stability with Multiple Freeze-Thaw Tolerance!
New Product Launch – Bovine Serum Albumin (BSA) for Clinical Biochemistry, Immunoassays, and Molecular Detection!
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HRP Blotting Dura ECL
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HRP Blotting Dura ECL is an enhanced, long-lasting chemiluminescent substrate ideal for quantitative Western blotting. It provides stable light output and is suitable for mid-femtogram level detection in Western blot analysis.
HRP Blotting Dura ECL
HRP Blotting Dura ECL is an enhanced, long-lasting chemiluminescent substrate ideal for quantitative Western blotting. It provides stable light output and is suitable for mid-femtogram level detection in Western blot analysis.
Art no: ECMD
Package size: 100mL, 1000 mL, bulk
Components:
Reagent A: Art no: ECMD-A, Enhanced Luminol Solution
Reagent B: Art no: ECMD-B, Stabilized Peroxide Solution
Shelf life: 3 years
Storage: 2-8℃ protected from light.
Application:
Western Blot: Use after 1:1 premixing
Characteristics:
① Sensitivity: Mid-femtogram level.
② Stability: Working solution remains stable for up to 24 hours; kit is stable for 1 year at room temperature.
③ Compatibility: Suitable for both nitrocellulose and PVDF membranes.
④ Extended Signal Duration: Signal persists for up to 24 hours.
⑤ Primary Antibody Dilution: 1:1,000–1:50,000(mass concentration: 0. 02–1.0 µg/mL).
⑥ Secondary Antibody Dilution: 1:50,000–1:250,000 (mass concentration: 4–20 ng/mL).
Instruction:
Western Blot Detection Using the Substrate:
1. Membrane Washing:
After protein transfer, remove the blot from the transfer apparatus and wash the membrane with deionized water for 5 minutes to remove residual transfer buffer.
2. Membrane Blocking:
Block the membrane with DuraBlot™ Western Blot Blocking Solution (e.g., 1× Casein Blocking Buffer, 1× Protein‑Free Blocking Buffer) at room temperature with shaking for 30–60 minutes, or alternatively, block statically overnight at 2–8 °C.
3. Primary Antibody Incubation:
Dilute the primary antibody to 0. 02–1.0 µg/mL (or according to the manufacturer’s recommended dilution) using primary antibody diluent (or blocking solution). Incubate with the membrane with shaking for 1 hour at room temperature, or overnight at 2–8 °C.
4. Membrane Washing:
Wash the membrane with washing buffer (TBST, PBST, or other buffer containing 0.05% Tween 20) for 10 minutes. Repeat the wash twice.
5. Enzyme‑Labeled Secondary Antibody Incubation:
Dilute the enzyme‑labeled secondary antibody to 4–20 ng/mL or 1:50,000–1: 250,000 using secondary antibody diluent. Incubate the blot with shaking for 30–60 minutes at room temperature.
6. Membrane Washing:
Wash the membrane 6 times with washing buffer, 5 minutes each wash, to thoroughly remove any unbound conjugated antibody.
7. Substrate Preparation:
Prepare the substrate working solution by mixing equal volumes of Substrate A (Enhanced Luminol Solution) and Stable Peroxide Solution B (e.g., 5 mL + 5 mL). Use sufficient volume to ensure complete wetting of the membrane without drying (0.1 mL/cm²).
8. Substrate Incubation:
Incubate the membrane with the substrate working solution for 5 minutes.
9. Membrane Imaging:
① Remove the blot from the working solution and place it in a plastic protector or clear plastic wrap.
② Remove excess liquid with lint‑free absorbent paper and carefully eliminate air bubbles from the membrane surface.
③ Image the blot using an imaging system.
Note: This product is highly sensitive for detection. Careful optimization of individual assay components (antibodies, conjugates, etc.) is essential to minimize nonspecific reactions and avoid high background.
For research use only or for further use in the production of detection kits. Not for human or animal use!
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