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0Enhanced ECL Chemiluminescent Substrate Kit
Product Description
Enhanced ECL Chemiluminescent Substrate Kit is designed to detect antibodies and associated antigens directly or indirectly labeled with horseradish peroxidase (HRP). The principle of Enhanced ECL Chemiluminescent Substrate Kit is that proteins or nucleic acids were transferred to the imprinted membrane after electrophoresis, and the target proteins on the membrane were bound by primary antibody and secondary antibody labeled with HRP, or the nucleic acids on the membrane were bound directly or indirectly by probes labeled with HRP. After washing the membrane, the ECL working solution prepared by the product was used to incubate the membrane at room temperature for several minutes. The imprinted membrane was wrapped with plastic wrap and fixed to the X-ray exposure Cassette. Then the X-ray film is pressed on the membrane in a darkroom and exposed for several seconds to several hours. After development and fixing, protein or nucleic acid bands can be clearly displayed on the X-ray film.
Product Components
|
Component |
36222ES60 (100 mL) |
36222ES76 (500 mL) |
|
|
36222-A |
Reagent A |
50 mL |
250 mL |
|
36222-B |
Reagent B |
50 mL |
250 mL |
Shipping and Storage
The products are shipped at ambient temperature and can be stored at room temperature for one year. If not used for a long time, it is recommended to store at 4℃ to extend the validity period.
[Notes]: Reagent A (36222-A) should be stored away from the light!
Instructions (X-ray film, for example)
1. Routine electrophoresis, transfer membrane, antibody labeled with HRP or nucleic acid probe labeled with HRP incubation and washing membrane.
[Notes]: ECL luminescent solution is the color substrate of HRP, so the detection system must be based on HRP enzyme-labeled antibody or nucleic acid probe.
2. At the same time as washing the film for the last time, the luminescent working solution was prepared fresh (100-200 μL luminescent working solution/cm2 membrane): take the same volume of Reagent A and B, mix well for using later.
[Notes]: Take Reagent A and Reagent B must use different tips. It is recommended to use the working fluid immediately. It can still be used after a few hours at room temperature, but the sensitivity is slightly reduced.
3. Use flat tweezers to take out the membrane, and put it on filter paper to drain the lotion, do not make the membrane completely dry. Completely immerse the membrane in luminescent working solution, with sufficient contact with luminescent working fluid. Incubate at room temperature for 1-2 minutes and prepare for tablet exposure immediately.
4. Pick up the membrane with tweezers and lay it on filter paper to drain the luminescent working fluid. But do not wash off the luminescent fluid.
5. Lay a piece of plastic wrap larger than the membrane on the inner surface of the X-ray film obscura. Attach the imprinted membrane to the plastic wrap, fold the plastic wrap completely around the imprinted membrane, and remove the gas bubble and fold, can cut off the edge of excess plastic wrap. Use filter paper to suck up excess luminous working fluid. Fix the plastic wrap covering the imprinted membrane in the obscura with tape, with the protein band facing upwards.
6. A piece of X-ray film was taken from the darkroom and placed on the wrapped membrane. The film was pressed and exposed for 30 secs-2 mins. Then the film was developed and fixed.
[Notes]: The exposure time should be adjusted according to the exposure intensity. If the background is too high, use two X-ray films at the same time.
Other exposure methods
If the use of CCD photography: the membrane can be placed in the working fluid, after the boot according to the use of the instructions, remove the film, and take photos. In addition, the measurement parameters of the machine can be adjusted according to the situation to improve the signal-to-noise ratio.
Cautions
1. For your safety and health, please wear lab coats and disposable gloves for the operation.
2. Bubbles should be avoided in membrane transfer, sealing, and incubation. In addition, wearing gloves will avoid leaving fingerprints on the membrane and keep it clean.
3. Prolonged exposure or excessive protein will deepen the background and make the change of band strength lose linear relation. Underexposure will blur the bands.
4. Some plastic wraps may quench the fluorescence when wrapping the imprinted membrane. High-quality plastic wrap should be selected.
5. Avoid placing multiple membranes in the same washing membrane box, as mutual adsorption or friction may cause a deep background.
6. The position and size of the bands on the film can be accurately determined using visible prestained protein markers and fluorescent-autoradiography exposure tags.
7. Use a biotin-avidin system and avoid using milk closure, which may cause the background to be too high.
8. Metal oxide particles may cause granular spots on the membrane. Avoid using rusty scissors and tweezers. Use plastic flat tweezers.
9. Sodium azide (NaN3) can inhibit HRP activity if the recovery of the HRP labeled probe or antibody should avoid using NaN3, if necessary, not more than 0.01%.
10. This product has no special toxicity, according to common chemical treatments.
11. For research use only!
[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] Li Y, Zhou S, Song H, Yu T, Zheng X, Chu Q. CaCO<sub>3</sub> 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)
[3] 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)
[4] 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)
[5] 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)
[6] 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)
[7] 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)
[8] 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. Published 2021 Jan 7. doi:10.1186/s13046-020-01816-3(IF:11.161)
[9] 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)
[10] 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)
[11] Song X, Gong Z, Liu K, Kou J, Liu B, Liu K. Baicalin combats glutamate excitotoxicity via protecting glutamine synthetase from ROS-induced 20S proteasomal degradation. Redox Biol. 2020;34:101559. doi:10.1016/j.redox.2020.101559(IF:9.986)
[12] Pan Y, He X, Li C, et al. Neuronal activity recruits the CRTC1/CREB axis to drive transcription-dependent autophagy for maintaining late-phase LTD. Cell Rep. 2021;36(3):109398. doi:10.1016/j.celrep.2021.109398(IF:9.423)
[13] Li KT, He X, Zhou G, et al. Rational designing of oscillatory rhythmicity for memory rescue in plasticity-impaired learning networks. Cell Rep. 2022;39(2):110678. doi:10.1016/j.celrep.2022.110678(IF:9.423)
[14] Zhang FL, Cao JL, Xie HY, et al. Cancer-Associated MORC2-Mutant M276I Regulates an hnRNPM-Mediated CD44 Splicing Switch to Promote Invasion and Metastasis in Triple-Negative Breast Cancer. Cancer Res. 2018;78(20):5780-5792. doi:10.1158/0008-5472.CAN-17-1394(IF:9.130)
[15] Sun R, Xie HY, Qian JX, et al. FBXO22 Possesses Both Protumorigenic and Antimetastatic Roles in Breast Cancer Progression. Cancer Res. 2018;78(18):5274-5286. doi:10.1158/0008-5472.CAN-17-3647(IF:9.130)
[16] Li M, Li A, Zhou S, Lv H, Yang W. SPAG5 upregulation contributes to enhanced c-MYC transcriptional activity via interaction with c-MYC binding protein in triple-negative breast cancer. J Hematol Oncol. 2019;12(1):14. Published 2019 Feb 8. doi:10.1186/s13045-019-0700-2(IF:8.731)
[17] Yin Q, Sun L, Cai X, et al. Lidocaine Ameliorates Psoriasis by Obstructing Pathogenic CGRP Signaling‒Mediated Sensory Neuron‒Dendritic Cell Communication. J Invest Dermatol. 2022;142(8):2173-2183.e6. doi:10.1016/j.jid.2022.01.002(IF:8.551)
[18] Chen W, Song J, Liu S, et al. USP9X promotes apoptosis in cholangiocarcinoma by modulation expression of KIF1Bβ via deubiquitinating EGLN3. J Biomed Sci. 2021;28(1):44. Published 2021 Jun 10. doi:10.1186/s12929-021-00738-2(IF:8.410)
[19] Lu J, Wang C, Cheng X, et al. A breakdown in microglial metabolic reprogramming causes internalization dysfunction of α-synuclein in a mouse model of Parkinson's disease. J Neuroinflammation. 2022;19(1):113. Published 2022 May 22. doi:10.1186/s12974-022-02484-0(IF:8.322)
[20] Yang YL, Zhang Y, Li DD, et al. RNF144A functions as a tumor suppressor in breast cancer through ubiquitin ligase activity-dependent regulation of stability and oncogenic functions of HSPA2. Cell Death Differ. 2020;27(3):1105-1118. doi:10.1038/s41418-019-0400-z(IF:8.086)
[21] Gao X, Liu S, Ding C, et al. Comparative effects of genistein and bisphenol A on non-alcoholic fatty liver disease in laying hens. Environ Pollut. 2021;288:117795. doi:10.1016/j.envpol.2021.117795(IF:8.071)
[22] Tan Y, Deng W, Zhang Y, et al. A marine fungus-derived nitrobenzoyl sesquiterpenoid suppresses receptor activator of NF-κB ligand-induced osteoclastogenesis and inflammatory bone destruction. Br J Pharmacol. 2020;177(18):4242-4260. doi:10.1111/bph.15179(IF:7.730)
[23] Lv Q, Xing Y, Liu Y, et al. Didymin switches M1-like toward M2-like macrophage to ameliorate ulcerative colitis via fatty acid oxidation. Pharmacol Res. 2021;169:105613. doi:10.1016/j.phrs.2021.105613(IF:7.658)
[24] Cheng B, Li X, Dai K, et al. Triggering Receptor Expressed on Myeloid Cells-2 (TREM2) Interacts With Colony-Stimulating Factor 1 Receptor (CSF1R) but Is Not Necessary for CSF1/CSF1R-Mediated Microglial Survival. Front Immunol. 2021;12:633796. Published 2021 Mar 25. doi:10.3389/fimmu.2021.633796(IF:7.561)
[25] Lv N, Jin S, Liang Z, et al. PP2Cδ Controls the Differentiation and Function of Dendritic Cells Through Regulating the NSD2/mTORC2/ACLY Pathway. Front Immunol. 2022;12:751409. Published 2022 Jan 7. doi:10.3389/fimmu.2021.751409(IF:7.561)
[26] Lv W, Yu H, Han M, et al. Analysis of Tumor Glycosylation Characteristics and Implications for Immune Checkpoint Inhibitor's Efficacy for Breast Cancer. Front Immunol. 2022;13:830158. Published 2022 Apr 4. doi:10.3389/fimmu.2022.830158(IF:7.561)
[27] Xu M, Li N, Fan X, et al. Differential Effects of Toll-Like Receptor Signaling on the Activation of Immune Responses in the Upper Respiratory Tract. Microbiol Spectr. 2022;10(1):e0114421. doi:10.1128/spectrum.01144-21(IF:7.171)
[28] Xie J, Yu J, Zhang Z, et al. AMPK pathway is implicated in low level lead-induced pubertal testicular damage via disordered glycolysis. Chemosphere. 2022;291(Pt 1):132819. doi:10.1016/j.chemosphere.2021.132819(IF:7.086)
[29] Lu X, Ji C, Jiang L, et al. Tumour microenvironment-based molecular profiling reveals ideal candidates for high-grade serous ovarian cancer immunotherapy. Cell Prolif. 2021;54(3):e12979. doi:10.1111/cpr.12979(IF:6.831)
[30] Wang Y, Wen Y, Xiao P, et al. Di-n-butyl phthalate promotes lipid accumulation via the miR200c-5p-ABCA1 pathway in THP-1 macrophages. Environ Pollut. 2020;264:114723. doi:10.1016/j.envpol.2020.114723(IF:6.793)
[31] Zhang Z, Huang Y, Xu N, et al. PLK1 Mitigates Intervertebral Disc Degeneration by Delaying Senescence of Nucleus Pulposus Cells. Front Cell Dev Biol. 2022;10:819262. Published 2022 Mar 14. doi:10.3389/fcell.2022.819262(IF:6.684)
[32] Lv W, Liu S, Zhang Q, Hu W, Wu Y, Ren Y. Circular RNA CircCOL5A1 Sponges the MiR-7-5p/Epac1 Axis to Promote the Progression of Keloids Through Regulating PI3K/Akt Signaling Pathway. Front Cell Dev Biol. 2021;9:626027. Published 2021 Jan 21. doi:10.3389/fcell.2021.626027(IF:6.684)
[33] Chen S, Zhang Z, Zhang B, et al. CircCDK14 Promotes Tumor Progression and Resists Ferroptosis in Glioma by Regulating PDGFRA. Int J Biol Sci. 2022;18(2):841-857. Published 2022 Jan 1. doi:10.7150/ijbs.66114(IF:6.582)
[34] Tao J, Qiu J, Lu L, et al. ZBTB20 Positively Regulates Oxidative Stress, Mitochondrial Fission, and Inflammatory Responses of ox-LDL-Induced Macrophages in Atherosclerosis. Oxid Med Cell Longev. 2021;2021:5590855. Published 2021 Mar 9. doi:10.1155/2021/5590855(IF:6.543)
[35] Cheng ST, Wang FF, Qian W. Cyclic-di-GMP binds to histidine kinase RavS to control RavS-RavR phosphotransfer and regulates the bacterial lifestyle transition between virulence and swimming. PLoS Pathog. 2019;15(8):e1007952. Published 2019 Aug 13. doi:10.1371/journal.ppat.1007952(IF:6.463)
[36] Liu J, Shen Z, Tang J, et al. Extracellular DNA released by glycine-auxotrophic Staphylococcus epidermidis small colony variant facilitates catheter-related infections. Commun Biol. 2021;4(1):904. Published 2021 Jul 22. doi:10.1038/s42003-021-02423-4(IF:6.268)
[37] Qiu L, Yang X, Wu J, Huang C, Miao Y, Fu Z. HIST2H2BF Potentiates the Propagation of Cancer Stem Cells via Notch Signaling to Promote Malignancy and Liver Metastasis in Colorectal Carcinoma. Front Oncol. 2021;11:677646. Published 2021 Aug 12. doi:10.3389/fonc.2021.677646(IF:6.244)
[38] Ding J, Zhang X, Chen C, Huang Y, Yu X, Li X. Ultra pH-sensitive polymeric nanovesicles co-deliver doxorubicin and navitoclax for synergetic therapy of endometrial carcinoma. Biomater Sci. 2020;8(8):2264-2273. doi:10.1039/d0bm00112k(IF:6.183)
[39] Zou Y, Xiao F, Song L, et al. A folate-targeted PEGylated cyclodextrin-based nanoformulation achieves co-delivery of docetaxel and siRNA for colorectal cancer. Int J Pharm. 2021;606:120888. doi:10.1016/j.ijpharm.2021.120888(IF:5.875)
[40] Zhenzhen L, Wenting L, Jianmin Z, et al. miR-146a-5p/TXNIP axis attenuates intestinal ischemia-reperfusion injury by inhibiting autophagy via the PRKAA/mTOR signaling pathway. Biochem Pharmacol. 2022;197:114839. doi:10.1016/j.bcp.2021.114839(IF:5.858)
[41] An L, Peng LY, Sun NY, et al. Tanshinone IIA Activates Nuclear Factor-Erythroid 2-Related Factor 2 to Restrain Pulmonary Fibrosis via Regulation of Redox Homeostasis and Glutaminolysis. Antioxid Redox Signal. 2019;30(15):1831-1848. doi:10.1089/ars.2018.7569(IF:5.828)
[42] Yang L, Cao N, Miao Y, Dai Y, Wei Z. Morin Acts as a USP7 Inhibitor to Hold Back the Migration of Rheumatoid Arthritis Fibroblast-Like Synoviocytes in a "Prickle1-mTORC2" Dependent Manner. Mol Nutr Food Res. 2021;65(19):e2100367. doi:10.1002/mnfr.202100367(IF:5.820)
[43] Pan B, Zheng L, Fang J, et al. Azilsartan Suppresses Osteoclastogenesis and Ameliorates Ovariectomy-Induced Osteoporosis by Inhibiting Reactive Oxygen Species Production and Activating Nrf2 Signaling. Front Pharmacol. 2021;12:774709. Published 2021 Nov 26. doi:10.3389/fphar.2021.774709(IF:5.811)
[44] Tan Y, Ke M, Li Z, et al. A Nitrobenzoyl Sesquiterpenoid Insulicolide A Prevents Osteoclast Formation via Suppressing c-Fos-NFATc1 Signaling Pathway. Front Pharmacol. 2022;12:753240. Published 2022 Jan 17. doi:10.3389/fphar.2021.753240(IF:5.811)
[45] Wang C, Huang W, Lu J, Chen H, Yu Z. TRPV1-Mediated Microglial Autophagy Attenuates Alzheimer's Disease-Associated Pathology and Cognitive Decline. Front Pharmacol. 2022;12:763866. Published 2022 Jan 18. doi:10.3389/fphar.2021.763866(IF:5.811)
[46] Yuan J, Li X, Fang N, et al. Perilla Leaf Extract (PLE) Attenuates COPD Airway Inflammation via the TLR4/Syk/PKC/NF-κB Pathway In Vivo and In Vitro. Front Pharmacol. 2022;12:763624. Published 2022 Jan 4. doi:10.3389/fphar.2021.763624(IF:5.811)
[47] Sa R, Guo M, Liu D, Guan F. AhR Antagonist Promotes Differentiation of Papillary Thyroid Cancer via Regulating circSH2B3/miR-4640-5P/IGF2BP2 Axis. Front Pharmacol. 2021;12:795386. Published 2021 Dec 23. doi:10.3389/fphar.2021.795386(IF:5.811)
[48] Qiao L, Fang L, Zhu J, et al. Total Flavone of Abelmoschus manihot Ameliorates TNBS-Induced Colonic Fibrosis by Regulating Th17/Treg Balance and Reducing Extracellular Matrix. Front Pharmacol. 2021;12:769793. Published 2021 Dec 23. doi:10.3389/fphar.2021.769793(IF:5.811)
[49] Yang H, Wang L, Zang C, et al. Squamosamide Derivative FLZ Diminishes Aberrant Mitochondrial Fission by Inhibiting Dynamin-Related Protein 1. Front Pharmacol. 2021;12:588003. Published 2021 Mar 19. doi:10.3389/fphar.2021.588003(IF:5.811)
[50] Tian X, Guo M, Zhang X, et al. Strongylocentrotus nudus Eggs Polysaccharide Enhances Macrophage Phagocytosis Against E.coli Infection by TLR4/STAT3 Axis. Front Pharmacol. 2022;13:807440. Published 2022 Mar 17. doi:10.3389/fphar.2022.807440(IF:5.811)
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