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methylflashtm methylated dna quantification kit  (EpiGentek)


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    Structured Review

    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
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    Images

    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

    Related Articles

    other:

    Article Title: Dynamic Changes in Oxidative Stress and Epigenetic Modifications in the Ventral Mesencephalon and Striatum of MPTP-Treated Mice: Implications for Parkinson's Disease Pathogenesis.
    Article Snippet: Quantification of 5‐Hydroxymethylcytosine Total 5-Hydroxymethylcytosine (5-hmC) content of genomic DNA was determined using the MethylFlash Hydroxymethylated DNA Quantification Kit (Colorimetric) (Epigentek Group Inc®, Farmingdale, USA Cat. No. P-1032).

    Article Title: Dynamic Changes in Oxidative Stress and Epigenetic Modifications in the Ventral Mesencephalon and Striatum of MPTP-Treated Mice: Implications for Parkinson's Disease Pathogenesis.
    Article Snippet: Quantification of 5‐Methylcytosine Total 5-Methylcytosine (5-mC) content in genomic DNA was measured using the MethylFlash Global DNA Methylation (5-mC) ELISA Easy Kit (Colorimetric) (Epigentek Group Inc®, Farmingdale, USA; Cat. No. P-1030).

    Article Title: Metabolic imprinting drives epithelial memory during mucosal fungal infection
    Article Snippet: Global DNA methylation levels were assessed using the MethylFlashTM Global DNA Methylation (5-mC) ELISA Easy Kit (EpigenTek, New York, USA), following the manufacturer’s protocols.

    Article Title: Chemotherapy treatment alters DNA methylation patterns in the prefrontal cortex of female rat brain.
    Article Snippet: 100ng of DNA from each sample was used to determine the global methylation level using Epigentek’s colorimetric MethylFlash Global DNA Methylation (5-mC) ELISA Easy Kit and MethylFlash Global DNA Hydroxy methylation (5-hmC) ELISA Easy Kit.

    Methylation:

    Article Title: Maternal Electronic Cigarette Exposure Induces Dysregulation of Autophagy via Oxidative Stress/DNA Methylation in Pulmonary Hypertension Offspring.
    Article Snippet: .. 5‐Methylcytosine (5‐mC) levels were quantified with a MethylFlashTM Methylated 5mC DNA Quantification Kit (EpigenTek, USA) according to the manufacturer’s instructions. ..

    Article Title: Phenotypic plasticity in Mediterranean gorgonians Eunicella singularis and Paramuricea clavata at high temperature and low pH.
    Article Snippet: • The oxygen consumption of the gorgonian corals increased at high

    DNA Methylation Assay:

    Article Title: Distribution patterns of N6-methyladenine in the rye genome
    Article Snippet: The DNA quality and quantity were assessed using 0.8% (w/v) agarose (Serva, Germany) gel electrophoresis in 1×TBE buffer and the spectrophotometric analysis (NanoDropTM 2000c; Thermo Fisher ScientificTM, USA). .. MethylFlashTM m 6 A DNA Methylation ELISA Kit (Epigentek, USA) was used to assess the global level of 6 mA in the rye genomic DNA. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Distribution patterns of N6-methyladenine in the rye genome
    Article Snippet: The DNA quality and quantity were assessed using 0.8% (w/v) agarose (Serva, Germany) gel electrophoresis in 1×TBE buffer and the spectrophotometric analysis (NanoDropTM 2000c; Thermo Fisher ScientificTM, USA). .. MethylFlashTM m 6 A DNA Methylation ELISA Kit (Epigentek, USA) was used to assess the global level of 6 mA in the rye genomic DNA. ..

    Immunoprecipitation:

    Article Title: Phenotypic plasticity in Mediterranean gorgonians Eunicella singularis and Paramuricea clavata at high temperature and low pH.
    Article Snippet: • The oxygen consumption of the gorgonian corals increased at high



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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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    Image Search Results


    Comparison of methylation data between MeDIP/NimbleGen Promoter + CpGi Array and Sequenom MassArray: Stk31 CpGi promoter region . The Stk31 gene region with a CpGi promoter region that is methylated in the liver, but not in ES or testis cells. The box indicates the position of the methylation peak in the liver that overlaps with the CpGi promoter region analyzed by Sequenom. In the epigram of Sequenom methylation analysis shown at the bottom panel, the blue circles indicate 100% methylation, green circles indicate around 50%methylation and yellow circles indicate 0% methylation. The results from biological duplicate samples are shown. The bottom lane (Sequenom Methylation Analysis) shows the results of analysis of testis DNA after in vitro methylation using M. Sss1 methylase (New England Biolabs).

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    Article Title: Genome-wide survey reveals dynamic widespread tissue-specific changes in DNA methylation during development

    doi: 10.1186/1471-2164-12-231

    Figure Lengend Snippet: Comparison of methylation data between MeDIP/NimbleGen Promoter + CpGi Array and Sequenom MassArray: Stk31 CpGi promoter region . The Stk31 gene region with a CpGi promoter region that is methylated in the liver, but not in ES or testis cells. The box indicates the position of the methylation peak in the liver that overlaps with the CpGi promoter region analyzed by Sequenom. In the epigram of Sequenom methylation analysis shown at the bottom panel, the blue circles indicate 100% methylation, green circles indicate around 50%methylation and yellow circles indicate 0% methylation. The results from biological duplicate samples are shown. The bottom lane (Sequenom Methylation Analysis) shows the results of analysis of testis DNA after in vitro methylation using M. Sss1 methylase (New England Biolabs).

    Article Snippet: Sequenom MassArray quantitative methylation analysis of bisulfite treated DNA was performed as an additional means to confirm DNA methylation peaks obtained by MeDIP/NimbleGen Array for both selected and randomly chosen MeDIP methylated regions.

    Techniques: Methylation, Methylated DNA Immunoprecipitation, In Vitro

    ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Article Snippet: TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells.

    Techniques: Expressing, DNA Methylation Assay, Sequencing, Comparison, Methylation

    DNA methylation analysis of chicken KLF7 in adipose tissues. a . The CpG density of the genomic region of chicken KLF7 analyzed by CpGplot (Version 6.6.0). b . The CpG loci analyzed in the promoter and Exon 2. c . The levels of DNA methylation of CpG loci analyzed. d . The comparison of DNA methylation levels between the promoter and Exon 2. Asterisks indicate a significant difference between the promoter and Exon 2 (Student’s t test, *** P < 0.001)

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: DNA methylation analysis of chicken KLF7 in adipose tissues. a . The CpG density of the genomic region of chicken KLF7 analyzed by CpGplot (Version 6.6.0). b . The CpG loci analyzed in the promoter and Exon 2. c . The levels of DNA methylation of CpG loci analyzed. d . The comparison of DNA methylation levels between the promoter and Exon 2. Asterisks indicate a significant difference between the promoter and Exon 2 (Student’s t test, *** P < 0.001)

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay, Comparison

    Datasets characteristics of DNA  methylation  of chicken KLF7 in abdominal adipose tissues

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Datasets characteristics of DNA methylation of chicken KLF7 in abdominal adipose tissues

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay

    Association analysis of DNA methylation with KLF7 transcripts and blood metabolic indicators. a . The r value of the Spearman correlation analysis. b . The P value of the Spearman correlation analysis. c . The number of samples used in the Spearman correlation analysis. d . The regression analysis between the DNA methylation of PCpG6 and KLF7 transcripts in chicken adipose tissue

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Association analysis of DNA methylation with KLF7 transcripts and blood metabolic indicators. a . The r value of the Spearman correlation analysis. b . The P value of the Spearman correlation analysis. c . The number of samples used in the Spearman correlation analysis. d . The regression analysis between the DNA methylation of PCpG6 and KLF7 transcripts in chicken adipose tissue

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay

    Schematic representation of the relationship of DNA methylation, KLF7 transcripts, glycaemia and abdominal fat content in chicken. The negative associations marked red and blue were confirmed by the reference [ , ], respectively

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Schematic representation of the relationship of DNA methylation, KLF7 transcripts, glycaemia and abdominal fat content in chicken. The negative associations marked red and blue were confirmed by the reference [ , ], respectively

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: DNA Methylation Assay

    Oligonucleotides used in the current study

    Journal: BMC Genetics

    Article Title: DNA methylation of CpG sites in the chicken KLF7 promoter and Exon 2 in association with mRNA expression in abdominal adipose tissue and blood metabolic indicators

    doi: 10.1186/s12863-020-00923-6

    Figure Lengend Snippet: Oligonucleotides used in the current study

    Article Snippet: The methylation level of individual units was measured by Quantitative Methylation Analysis (MassARRAY EpiTYOER, Sequenom, San Diego, CA).

    Techniques: Methylation

    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