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Description
The Enhanced ECL Chemiluminescent Substrate Kit is designed for the detection of horseradish peroxidase (HRP)-labeled primary or secondary antibodies and their associated antigens. The principle involves separating proteins or nucleic acids by electrophoresis, transferring them onto a blotting membrane, and then incubating with a primary antibody followed by an HRP-conjugated secondary antibody, or using an HRP-labeled probe to directly or indirectly bind target nucleic acids on the membrane. After washing, the membrane is incubated at room temperature for several minutes with freshly prepared ECL working solution from this kit. The membrane is then wrapped in plastic wrap, placed face-up into an X-ray film cassette, and exposed to X-ray film in a darkroom for seconds to hours. Following development and fixation, protein or nucleic acid bands are clearly visualized on the X-ray film.
This kit features a unique luminescent substrate system that reduces background signal while incorporating a novel oxidant to significantly enhance stability, allowing the reagent to remain stable for one year at room temperature. In addition to X-ray film detection, the chemiluminescent signal can be directly captured using a fluorescence CCD imager. This product is primarily intended for Western blot (WB) applications and chemiluminescent immunoassay systems.
Features
Superb Sensitivity: Detects antigens at picogram to low femtogram levels for ultra-sensitive Western blot results.
High Signal-to-Noise Ratio: Advanced luminescent substrates deliver strong signals with minimal background noise.
Antibody-Efficient: Optimized formulation enhances antibody binding, reducing antibody consumption.
Excellent Value: Delivers top-tier performance at a competitive price.
Outstanding Stability: Improved oxidant system ensures stable storage at 4°C for up to 12 months.
Specifications
|
Applicable Samples |
Target with high abundance, routine protein samples |
|
Detection Sensitivity |
Medium picogram level |
|
Signal Duration |
< 2 h |
|
Applicable Detection System |
All reagents are suitable for X-ray film, CCD imaging devices |
|
Primary Antibody Dilution Concentration |
1:1000-1:4000 |
|
Secondary Antibody Dilution Concentration |
1:1000-1:4000 |
|
Recommended Membrane |
NC membrane |
|
Storage Stability |
Room temperature for 1 year |
|
Advantages |
Strongest signal among the same level of sensitivity |
Components
|
Components No. |
Name |
36222ES60 |
36222ES76 |
|
36222-A |
Solution A |
50 mL |
250 mL |
|
36222-B |
Solution B |
50 mL |
250 mL |
Storage
For frequent use, store at room temperature where it remains stable for up to one year.
For long-term storage, storing at 4 °C is recommended to extend valid life.
[Note]: Solution A (36222-A) must be protected from light.
Application
Chemiluminescence ELISA
Western Blot
Dot Blot-DNA/RNA
Southern blot-DNA
Northern blot-RNA
FAQ
Q: What is the dilution concentration of the primary antibody used in this product?
A: 1:1000 - 1:4000.
Q: What is the dilution concentration of the secondary antibody used in this product?
A: 1:1000 - 1:4000.
Q: What are the differences between this product and 36208?
A: This product has a high target abundance and a short detection time, making it suitable for routine protein samples. 36208 belongs to a highly sensitive colorimetric kit, with low target abundance, limited samples, and a longer detection time.
Documents
Safety Data Sheet
Manuals
36222_Manual_Ver.EN20250923.pdf
Rerference
1. He X, Li J, Zhou G, et al. Gating of hippocampal rhythms and memory by synaptic plasticity in inhibitory interneurons. Neuron. 2021;109(6):1013-1028.e9. doi:10.1016/j.neuron.2021.01.014. IF = 17.173
2. Wang X, Zhang S, Jin D, et al. μ-opioid receptor agonist facilitates circulating tumor cell formation in bladder cancer via the MOR/AKT/Slug pathway: a comprehensive study including randomized controlled trial. Cancer Commun (Lond). 2023;43(3):365-386. doi:10.1002/cac2.12408. IF = 16.2
3. Li Y, Zhou S, Song H, Yu T, Zheng X, Chu Q. CaCO₃ nanoparticles incorporated with KAE to enable amplified calcium overload cancer therapy. Biomaterials. 2021;277:121080. doi:10.1016/j.biomaterials.2021.121080. IF = 12.479
4. Xu Z, Xu J, Sun S, et al. Mecheliolide elicits ROS-mediated ERS-driven immunogenic cell death in hepatocellular carcinoma. Redox Biol. 2022;54:102351. doi:10.1016/j.redox.2022.102351. IF = 11.799
5. Cai Z, Zhang Y, Zhang W, et al. Arsenic retention in erythrocytes and excessive erythrophagocytosis is related to low selenium status by impaired redox homeostasis. Redox Biol. 2022;52:102321. doi:10.1016/j.redox.2022.102321. IF = 11.799
6. Kaushik AC, Wu Q, Lin L, et al. Exosomal ncRNAs profiling of mycobacterial infection identified miRNA-185-5p as a novel biomarker for tuberculosis. Brief Bioinform. 2021;22(6):bbab210. doi:10.1093/bib/bbab210. IF = 11.622
7. Hu P, Li H, Sun W, et al. Cholesterol-associated lysosomal disorder triggers cell death of hematological malignancy: dynamic analysis on cytotoxic effects of LW-218. Acta Pharm Sin B. 2021;11(10):3178-3192. doi:10.1016/j.apsb.2021.02.004. IF = 11.614
8. Wang X, Ni J, You Y, et al. SNX10-mediated LPS sensing causes intestinal barrier dysfunction via a caspase-5-dependent signaling cascade. EMBO J. 2021;40(24):e108080. doi:10.15252/embj.2021108080. IF = 11.598
9. Jiang K, Chen H, Fang Y, et al. Exosomal ANGPTL1 attenuates colorectal cancer liver metastasis by regulating Kupffer cell secretion pattern and impeding MMP9-induced vascular leakiness. J Exp Clin Cancer Res. 2021;40(1):21. doi:10.1186/s13046-020-01816-3. IF = 11.161
10. Zhang S, Yang Z, Bao W, et al. SNX10 (sorting nexin 10) inhibits colorectal cancer initiation and progression by controlling autophagic degradation of SRC. Autophagy. 2020;16(4):735-749. doi:10.1080/15548627.2019.1632122. IF = 11.059
11. Liu L, Liang L, Yang C, Zhou Y, Chen Y. Extracellular vesicles of Fusobacterium nucleatum compromise intestinal barrier through targeting RIPK1-mediated cell death pathway. Gut Microbes. 2021;13(1):1-20. doi:10.1080/19490976.2021.1902718. IF = 10.245
Enhanced ECL substrates contain modified chemiluminescent enhancers that increase photon emission rate and prolong signal duration compared to standard ECL. This results in a brighter, more stable signal compatible with digital imaging systems and film-based detection, allowing detection of low-abundance targets with shorter exposure times and reduced background from non-specific binding.
Optimal antibody concentration depends on antibody affinity, target abundance, and membrane type. As a starting point, HRP-conjugated secondary antibodies are typically used at 1:5,000–1:20,000 dilution with Enhanced ECL. Titrate the antibody concentration empirically — excess HRP enzyme can increase background chemiluminescence, reducing signal-to-noise. Refer to the antibody manufacturer's datasheet for application-specific guidance.
Signal half-life varies by substrate formulation. Enhanced ECL substrates typically provide a signal window of 5–30 minutes, with peak intensity in the first 5–10 minutes. Capture images promptly after substrate application. For low-abundance targets requiring long exposures (>5 minutes), a longer-lasting Ultra or Pico ECL variant may be more appropriate.
Yes. This Enhanced ECL kit is validated for use with both PVDF (polyvinylidene fluoride) and nitrocellulose membranes. PVDF membranes generally give lower background and are preferred for stripping and re-probing. Ensure complete blocking (5% non-fat dry milk or BSA) prior to antibody incubation to minimize non-specific signal.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
