Magic Red – Quantification and monitoring of intracellular catechin-B activity over time in vitro. The small portion of Magic Red in this test is red after being separated from the active catechin. Analyze the fluorescent signal using fluorescence microscopy or a fluorescent plate reader.

Elevation of catechin enzyme in serum or extracellular matrix can indicate several pathologies. ICT’s Magic Red assay kit is used by researchers who attempt to quantify and monitor catecholamine activity in cultured cells and tissues. These test kits detect the activity of catechol-B To use Red Magic, add the component directly to the media, activate, and analyze. Because it is cell-permeable, it easily penetrates cell membranes and the membranes of internal organelles – no lysis or permeabilization processes are required. If the catechol protein is active, the magic red substrate is cleaved and the cresyl violet fluorophore fluoresces when excited. As the enzyme activity progresses and a small amount of Magic Red is broken off, the signal will grow, allowing the researcher to see the color develop over time. Samples can be analyzed by fluorescence microscopy or fluorescence plate readers. Hoechst 33342 and acridine orange are included in the kit to detect nuclear structure and lysosomal organelle formation.

Magic Red

Magic Red

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Lee, Y; Zhu, B.; Ren, X.; My, L.; Zhu, Y; Zhao, Y; Yang, C.; Yuan, C; Lee, H.; Tonga, X; Wang, Y; Du, J. CISD2 promotes ferritinophagy-mediated ferritin trafficking and ferroptosis through regulation of the p62-Keap1-NRF2 pathway. Cellular and Molecular Biology Letters 2022 September 30; doi:10.1186/s11658-022-00383-z Full abstract.

No, H.; Shen, K.; Meireles, M.; Talbot, W. The lysosomal regulatory system for microglia development and function Science Advances 2022 September 2; doi:10.1126/sciadv.abp8321 Details

Akter, E.; Tasaki, Y; Mori, Y; Nakai, K; Hachiya, K; Lynn, H.; Kano, M.; Kamasaki, T; Tanabe, K; Umeda, Y; Yamano, S.; Fujita, Y; Cone, Non-defective autophagic vacuoles are essential for cellular competition. Room reports 2022 August 30; doi:10.1016/j.celrep.2022.111292 Article

Fernandez, MA; Baha, F.; Mother, L; Lee, Y; Schmidt, M.; Welch, E.; Moore, M.; Young-Pearce, TL Loss of the endosomal exchanger NHE6 leads to pathological changes in tau in human neurons. Stem Cell Reports 2022 Aug 17; doi:10.1016/j.stemcr.2022.08.001 Full report

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Reto, E.; Chowdhuri, SR; Siefert, NS; Schneider, M.; Whitman, M; Helm, D.; Pama, W. Nature Communications 2022 Aug 17; doi:10.1038/s41467-022-32515-6 Article

O, Y; Black, K; Yoshimura, SI; Hara, N.; Hasegawa, J.; Terawaki, S.; Kimura, Y; Ikenaka, K; Suitsugu, S.; Mochizuki, H.; It is important for amphisome-lysosome fusion during basal autophagy and subcellular autophagy. PLoS Genetics 2022 June 1; doi:10.1371/journal.pgen.1010264 Text

I am, M; Lee, P.; Wang, C.; Feng, X; As, Q. Chen, W.; Marty, M.; Zhang, W; Gao, C.; RI; Xu, H. Parkinson’s disease protein TMEM175 is a protein-activated proton channel in lysosomes. Room 2022 Jun 23; doi:10.1016/j.cell.2022.05.021 Full text

Magic Red

Now, T; Zhang, P.; Bailey, C.; Wang, Y; Itiprasert, W.; Davenport, MI; Zheng, P.; Liu, Y. Genes and Diseases 2022 May 1; doi:10.1016/j.gendis.2022.05.004 Full text

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Jong, W; Rawo, J.; And, J.; Sun, Y; Oh, hey; Wang, P.; Yes, Y; Bottom, X; Tanga, W.; Chen, J.; Zhou, H.; Wanga, X. Activation of STING signaling in chronic liver inflammation promotes mitochondrial DNA cytosolic leakage in aged macrophages. Nursing Home 2022 May 22; doi:10.1111/jcpe.13647 Abstract

Browning, CL; Green, A.; Gray, EP; Losing, R.; Kane, AB. Manganese dioxide nanoparticles induce mitochondrial toxicity in fish epithelial cells. Nanotoxicology 2021 Jan 27; 1-18 doi:10.1080/17435390.2021.1874562 Article

Lan J, Luo H, Wu R, Wang J, Zhou B, Zhang Y, Jiang Y, Zhu J. Arteriosclerosis of Thromb Vasc Biol Dec 2020; 40(12):2922-2940 doi: 10.1161/ATVBAHA.120.315151 Epub 2020 Oct 1. Full text.

Jager E, Murty S, Schmidt C, Hahn M, Strobel S, Peters A, Staubert C, Sungur P, Shukr T, Geisler M, Radusheva V, Raps S, Rothe K, Scholz R, Jung S, Wagner S, Pierre M . , Seifert O, Chang W, Estrella-Lopez I, Raulien N, Krone K, Strater N, Hoppener S, Schoenberg T, Rossol M, Wagner U. Nat Commun 2020 Aug 25; 11(1): 4243 doi:10.1038/s41467-020-17749-6 Full text

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Yin Q, Jian Y, Xu M, Huang X, Wang N, Liu Z, Li Q, Li J, Zhou H, Xu L, Wang Y, Yang C. CDK4/6 regulates lysosome biogenesis through TFEB/TFE3. J Cell Biol 2020 Aug 3; 219(8):e201911036 doi:10.1083/jcb.201911036 Full text

[ Abstract ] Almakelas E, Pelletier J, Tsiku C, Ambrosio S, Tauler A, Mauvezin C. Lysosomal degradation ensures proper chromosomal differentiation to prevent chromosomal instability. Autophagy 2020 Jun 23; 1-18 doi:10.1080/15548627.2020.1764727 Online ahead of print. All words

Song SB, Hwang ES. Increased ROS impairs lysosomal acidity and autophagy flux in glucose-deprived fibroblasts by activating the ATM and Erk pathways. Biomolecules 2020 May 13; 10(5):761 doi:10.3390/biom10050761 abstract

Magic Red

He M, Zhang T, Xu J, Qin S, Wang H, Zhao L, Wang X, Hu J, Wen J, Sai H, Jin Q, Guo Q, Lin L, Zhou B, Zhang H, Jia G, Wang C LSD1 promotes oocyte death by regulating the transcription of the autophagy adapter SQSTM1/p62. Aging Center March 2020; 19(3):e13102 doi: 10.1111/acel.13102 Epub 2020 Feb 19. Full text.

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Kratmer C, Levy M. Targeted delivery of aristatin-modified toxin in pancreatic cancer using aptamers. Molecular Therapy-Nucleic Acids 2017. 10:227-236 March 2018. doi.org/10.1016/j.omtn.2017.11.013. All words

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Magic Red

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Fujita N, Huang W, Lin TH, Groulx JF, Zhan S, Nguyen J, Kuchitou Y, Koyama-Honda I, Mizushima N, Fukuda M, Kiger A. Genetic screen in Drosophila tissue identifies a role for Rab2 in autophagy-mediated T -tubule remodeling and autophagy. ELIF 2017. Full text

Question: If you dilute the stock to 1/10 in H2O and aliquot for future use, how long can it be stored? I find that a large amount of stock in DMSO can be stored for 6 months

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Answer: When the stock solution is diluted 1:10 in diH2O, we recommend using it within 15 minutes to avoid hydrolysis of the substrate of Magic Red. Non-specific hydrolysis leads to unspecific fluorescence and enzymatic cleavage of the substrate. We do not recommend aliquoting or storing the diH2O-diluted substrate solution for future use. Yes, that’s right DMSO-reconstituted Magic Red stock can be stored protected from light for 6 months and can be diluted twice as much in that time.

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