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methylflashtm methylated dna quantification kit  (EpiGentek)


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    EpiGentek methylflashtm methylated dna quantification kit
    A The B16F10 melanoma tumor volume in each group was determined at the indicated time points. B Images of tumor nodules at the endpoint of the experiment. C The weight of the tumor nodules from each group at the endpoint of the experiment. D Western blot analysis of MBD2 in each group. E , F RT-PCR analysis of Cdh1 ( E ) and Cdh2 ( F ) in the indicated groups. G Representative images of E-cadherin and N-cadherin coimmunostaining in the tumor nodule sections. H Images of pulmonary metastatic nodules from the indicated groups. I Representative H&E staining of pulmonary metastatic nodules from the indicated groups. J MBD2 binds to <t>methylated</t> CpG <t>DNA</t> within the DDB2 promoter and thus represses DDB2 expression, resulting in tumor metastasis. Intravenous administration of liposomes carrying MBD2 siRNA suppressed tumor growth in an animal model bearing tumor metastasis. L-Scr siRNA: liposome-based Scrambled siRNA; L- MBD2 siRNA: liposome-based MBD2 siRNA. The data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.
    Methylflashtm Methylated Dna Quantification Kit, supplied by EpiGentek, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/massarray+quantitative+methylation+analysis+platform/pmc10160113-75-7-12?v=EpiGentek
    Average 90 stars, based on 1 article reviews
    methylflashtm methylated dna quantification kit - by Bioz Stars, 2026-07
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    1) Product Images from "MBD2 facilitates tumor metastasis by mitigating DDB2 expression"

    Article Title: MBD2 facilitates tumor metastasis by mitigating DDB2 expression

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-023-05804-1

    A The B16F10 melanoma tumor volume in each group was determined at the indicated time points. B Images of tumor nodules at the endpoint of the experiment. C The weight of the tumor nodules from each group at the endpoint of the experiment. D Western blot analysis of MBD2 in each group. E , F RT-PCR analysis of Cdh1 ( E ) and Cdh2 ( F ) in the indicated groups. G Representative images of E-cadherin and N-cadherin coimmunostaining in the tumor nodule sections. H Images of pulmonary metastatic nodules from the indicated groups. I Representative H&E staining of pulmonary metastatic nodules from the indicated groups. J MBD2 binds to methylated CpG DNA within the DDB2 promoter and thus represses DDB2 expression, resulting in tumor metastasis. Intravenous administration of liposomes carrying MBD2 siRNA suppressed tumor growth in an animal model bearing tumor metastasis. L-Scr siRNA: liposome-based Scrambled siRNA; L- MBD2 siRNA: liposome-based MBD2 siRNA. The data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.
    Figure Legend Snippet: A The B16F10 melanoma tumor volume in each group was determined at the indicated time points. B Images of tumor nodules at the endpoint of the experiment. C The weight of the tumor nodules from each group at the endpoint of the experiment. D Western blot analysis of MBD2 in each group. E , F RT-PCR analysis of Cdh1 ( E ) and Cdh2 ( F ) in the indicated groups. G Representative images of E-cadherin and N-cadherin coimmunostaining in the tumor nodule sections. H Images of pulmonary metastatic nodules from the indicated groups. I Representative H&E staining of pulmonary metastatic nodules from the indicated groups. J MBD2 binds to methylated CpG DNA within the DDB2 promoter and thus represses DDB2 expression, resulting in tumor metastasis. Intravenous administration of liposomes carrying MBD2 siRNA suppressed tumor growth in an animal model bearing tumor metastasis. L-Scr siRNA: liposome-based Scrambled siRNA; L- MBD2 siRNA: liposome-based MBD2 siRNA. The data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Techniques Used: Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Methylation, Expressing, Liposomes, Animal Model



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    During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through <t>methylation-dependent</t> transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom <t>MassARRAY</t> assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).
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    During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through methylation-dependent transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom MassARRAY assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).

    Journal: Chinese Journal of Cancer Research

    Article Title: Stomatin plays a suppressor role in non-small cell lung cancer metastasis

    doi: 10.21147/j.issn.1000-9604.2019.06.09

    Figure Lengend Snippet: During transforming growth factor β1 (TGFβ1)-induced epithelial-to-mesenchymal transition (EMT), stomatin was downregulated through methylation-dependent transcription mechanism. (A) Real-time polymerase chain reaction (PCR) was performed to validate the transcriptional change of stomatin identified in the microarray. **, P<0.001; (B) Two CpG islands spanning from −768 to −223 bp and from −143 to +122 bp upstream of STOM gene were identified by EMBOSS cpgplot program (upper). Schematic representation of −768/−223 and −143/+122 (marked with the grey square) CpG islands in the upstream of stomatin gene (lower). Red box marked the exon. The translation start site was position +1; (C) Measuring stomatin promoter methylation levels by Sequenom MassARRAY assay; (D) Sequenom MassARRAY analysis of methylation percentages of CpG sites at the stomatin promoter. The top panel showed the methylation percentages of 30 sites in human bronchial epithelial (HBE) and A549 cells. Dots with different color depths represent different methylation status. Dark dots represent high degrees of methylation. Light dots represent low degrees of methylation. The methylation percentages of 30 CpG sites were compared by two-tailed paired Student’s t test. In line graph, each dot represents an individual CpG dinucleotide. Green dots represent CpG dinucleotides in HBE cells. Red dots represent CpG sites in A549 cells. The corresponding detection sites were connected in a straight line. Real-time PCR was performed to evaluate the expression of stomatin in HBE and A549 cells (P<0.01); (E) The same sequenom MassArray and statistical analyses were also implemented to assess the methylation status of the indicated promoter region of stomatin in A549 and A549-5Aza cells (P<0.01); (F) A549 cells were exposed to TGFβ1 (5 ng/mL) for 2 d or 5 d with or without 5-Aza. Expression of DNMT3A and stomatin was analyzed by western blotting. β-actin was used as a loading control; (G) Sequenom MassArray analysis was implemented to assess the stomatin promoter methylation levels in A549, A549-TGFβ1, and A549-TGFβ1/5-Aza cells (P=0.005, P=0.004, respectively).

    Article Snippet: The methylation status of Stomatin promoter was detected using Sequenom MassARRAY quantitative methylation analysis platform (BGI tech, Shenzhen, China).

    Techniques: Methylation, Real-time Polymerase Chain Reaction, Microarray, Sequenom Massarray Assay, Two Tailed Test, Expressing, Western Blot, Control