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    EpiGentek antibody against histone h4k20me3
    The level of <t>H4K20me3</t> in cell cycle phases. H4K20me3 in ( A ) G1, ( B ) S, and ( C ) G2 phases of the cell cycle. G1 and G2 phases were determined according to the nuclear distribution pattern of H3S10 phosphorylation: the level of H3S10p is low in the G1 phase characterized by an appearance of tiny H3S10p-positive signals. G2 phase is characterized by robust H3S10p-positive signals in the cell nucleus. The S phase was recognized according to the distribution pattern of the mCherry-tagged PCNA protein that appeared at DNA lesions in the late S phase. The highest level of H4K20me3 at DNA lesions was in the G1 phase of the cell cycle, while cells in S and G2 phases of the cell cycle were characterized by reduced H4K20me3 at micro-irradiated chromatin. Scale bars represent 10 µm. ( D ) The phenomenon observed in panels A-C was confirmed by the use of the HeLa-Fucci cellular system showing G1 cells expressing RFP-cdt1 and G2 cells with GFP-geminin positivity. The cells in early S phase were subtly positive for both RFP-cdt1 and GFP-geminin. ( E ) The level of H4K20me3 (red) in mouse fibroblast over-expressing GFP-tagged BRCA1 protein (green). Scale bars in all panels represent 10 µm.
    Antibody Against Histone H4k20me3, supplied by EpiGentek, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The level of H4K20me3 in cell cycle phases. H4K20me3 in ( A ) G1, ( B ) S, and ( C ) G2 phases of the cell cycle. G1 and G2 phases were determined according to the nuclear distribution pattern of H3S10 phosphorylation: the level of H3S10p is low in the G1 phase characterized by an appearance of tiny H3S10p-positive signals. G2 phase is characterized by robust H3S10p-positive signals in the cell nucleus. The S phase was recognized according to the distribution pattern of the mCherry-tagged PCNA protein that appeared at DNA lesions in the late S phase. The highest level of H4K20me3 at DNA lesions was in the G1 phase of the cell cycle, while cells in S and G2 phases of the cell cycle were characterized by reduced H4K20me3 at micro-irradiated chromatin. Scale bars represent 10 µm. ( D ) The phenomenon observed in panels A-C was confirmed by the use of the HeLa-Fucci cellular system showing G1 cells expressing RFP-cdt1 and G2 cells with GFP-geminin positivity. The cells in early S phase were subtly positive for both RFP-cdt1 and GFP-geminin. ( E ) The level of H4K20me3 (red) in mouse fibroblast over-expressing GFP-tagged BRCA1 protein (green). Scale bars in all panels represent 10 µm.

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: The level of H4K20me3 in cell cycle phases. H4K20me3 in ( A ) G1, ( B ) S, and ( C ) G2 phases of the cell cycle. G1 and G2 phases were determined according to the nuclear distribution pattern of H3S10 phosphorylation: the level of H3S10p is low in the G1 phase characterized by an appearance of tiny H3S10p-positive signals. G2 phase is characterized by robust H3S10p-positive signals in the cell nucleus. The S phase was recognized according to the distribution pattern of the mCherry-tagged PCNA protein that appeared at DNA lesions in the late S phase. The highest level of H4K20me3 at DNA lesions was in the G1 phase of the cell cycle, while cells in S and G2 phases of the cell cycle were characterized by reduced H4K20me3 at micro-irradiated chromatin. Scale bars represent 10 µm. ( D ) The phenomenon observed in panels A-C was confirmed by the use of the HeLa-Fucci cellular system showing G1 cells expressing RFP-cdt1 and G2 cells with GFP-geminin positivity. The cells in early S phase were subtly positive for both RFP-cdt1 and GFP-geminin. ( E ) The level of H4K20me3 (red) in mouse fibroblast over-expressing GFP-tagged BRCA1 protein (green). Scale bars in all panels represent 10 µm.

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Irradiation, Expressing

    Schematic illustration of protein levels in DNA lesions in wild-type and Suv39h1/h2 dn mouse embryonic fibroblasts. Colored dots illustrate the levels of selected proteins at UVA-irradiated chromatin. The illustration demonstrates the appearance of the following proteins at DNA lesions: HP1β (pink circles), H3K9me3 (orange circles), H3K9ac (dark green circles), H4K20me3 (red circles), γH2AX (pale green circles), and 53BP1 (white circles). The selected micro-irradiated ROI is shown by a white rectangle. The figure represents an illustration of our results.

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: Schematic illustration of protein levels in DNA lesions in wild-type and Suv39h1/h2 dn mouse embryonic fibroblasts. Colored dots illustrate the levels of selected proteins at UVA-irradiated chromatin. The illustration demonstrates the appearance of the following proteins at DNA lesions: HP1β (pink circles), H3K9me3 (orange circles), H3K9ac (dark green circles), H4K20me3 (red circles), γH2AX (pale green circles), and 53BP1 (white circles). The selected micro-irradiated ROI is shown by a white rectangle. The figure represents an illustration of our results.

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Irradiation

    An interaction between 53BP1-H4K20me2/me3. ( A ) Immunoprecipitation (IP) experiments showed an interaction between H3K9me3 and 53BP1 or H4K20me2 and 53BP1 or H4K20me3 and the 53BP1 protein. HP1β protein did not interact with the 53BP1 protein. ( B ) Quantification of IP fragments from panel (A) studied in non-irradiated and γ-irradiated Suv39h1/h2 wt and Suv39h1/h2 dn cells. Asterisks (*) indicate statistical significance at P≤0.05.

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: An interaction between 53BP1-H4K20me2/me3. ( A ) Immunoprecipitation (IP) experiments showed an interaction between H3K9me3 and 53BP1 or H4K20me2 and 53BP1 or H4K20me3 and the 53BP1 protein. HP1β protein did not interact with the 53BP1 protein. ( B ) Quantification of IP fragments from panel (A) studied in non-irradiated and γ-irradiated Suv39h1/h2 wt and Suv39h1/h2 dn cells. Asterisks (*) indicate statistical significance at P≤0.05.

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Immunoprecipitation, Irradiation

    Quantification of western blot data on selected histone markers in non-irradiated and γ-irradiated cells. Using ImageJ and ImageQuant TL software, the levels of the following proteins (originated from Fig. 1A, B ) were quantified: H3K9me1, H3K9me2, H3K9me3, H3K9ac, H4K20me2, H4K20me3, H4K20ac, γH2AX, HP1β, 53BP1, and MDC1. The levels of modified histones were normalized to those of total H3 histones, and those of DDR-related proteins were normalized to those of α-tubulin. Quantification was performed in the following samples: ( A ) HAP1 wt and HAP1 mutant cells and ( B ) wt MEFs and Suv39h1/h2-deficient fibroblasts (MEFs).

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: Quantification of western blot data on selected histone markers in non-irradiated and γ-irradiated cells. Using ImageJ and ImageQuant TL software, the levels of the following proteins (originated from Fig. 1A, B ) were quantified: H3K9me1, H3K9me2, H3K9me3, H3K9ac, H4K20me2, H4K20me3, H4K20ac, γH2AX, HP1β, 53BP1, and MDC1. The levels of modified histones were normalized to those of total H3 histones, and those of DDR-related proteins were normalized to those of α-tubulin. Quantification was performed in the following samples: ( A ) HAP1 wt and HAP1 mutant cells and ( B ) wt MEFs and Suv39h1/h2-deficient fibroblasts (MEFs).

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Western Blot, Irradiation, Software, Modification, Mutagenesis

    The nuclear distribution pattern of H4K20me1/me2/me3 at DNA lesions. ( A ) The levels of ( a ) H4K20me1 (red) in γH2AX-positive DNA lesions (magenta), ( b ) H4K20me2 (red) in γH2AX-positive DNA lesions (magenta), and ( c ) H4K20me3 (red) in DNA lesions studied in parallel with γH2AX (magenta). ( B ) The level of ( a ) H4K20me3 and ( b ) 53BP1 in micro-irradiated ROI of the cells over-expressing JMJD2b histone demethylase, tagged by GFP. Panels Bb-aa show low level of 53BP1 at micro-irradiation induced DNA lesions in cells over-expressing GFP-tagged JMJD2b and panel Bb-bb documents 53BP1 recruitment to DSB sites in the cells with a normal expression of JMJD2b. Scale bars in all panels represent 5 µm.

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: The nuclear distribution pattern of H4K20me1/me2/me3 at DNA lesions. ( A ) The levels of ( a ) H4K20me1 (red) in γH2AX-positive DNA lesions (magenta), ( b ) H4K20me2 (red) in γH2AX-positive DNA lesions (magenta), and ( c ) H4K20me3 (red) in DNA lesions studied in parallel with γH2AX (magenta). ( B ) The level of ( a ) H4K20me3 and ( b ) 53BP1 in micro-irradiated ROI of the cells over-expressing JMJD2b histone demethylase, tagged by GFP. Panels Bb-aa show low level of 53BP1 at micro-irradiation induced DNA lesions in cells over-expressing GFP-tagged JMJD2b and panel Bb-bb documents 53BP1 recruitment to DSB sites in the cells with a normal expression of JMJD2b. Scale bars in all panels represent 5 µm.

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Irradiation, Expressing

    Histone signature in non-irradiated and irradiated cells without and with mutation or deletion in Suv39h1/h2 histone methyltransferases. Western blot analysis of γH2AX, H3K9me1, H3K9me2, H3K9me3, H3K9ac, H4K20me2, H4K20me3 and H4K20 acetylation. The levels of modified histones were normalized to that of total H3 histones. As DNA damage markers, 53BP1, MDC1 proteins, and the HP1β protein were studied and normalized to the level of α-tubulin. Protein levels were studied in ( A ) HAP1 wt and HAP1 cells with the mutation in the SUV39H1 gene and ( B ) in wt MEFs and Suv39h1/h2-deficient fibroblasts (MEFs). Non-irradiated cells and cells irradiated by 5 Gy of γ-rays (harvested 30 min and 24 hours after irradiation) were analyzed. ( C ) Cell cycle profiles were studied by flow cytometry in non-irradiated and γ-irradiated wt MEFs and non-irradiated and γ-irradiated Suv39h1/h2 dn mouse embryonic fibroblasts. Panel ( a ) shows a 30-min interval, and panel ( b ) shows a 24-h interval when the cells were harvested after irradiation (and related control samples). Using Mod-Fit software, the percentage of cells in G1 (red peak), S (dash blue peak) and G2-M (green peak) cell cycle phases was calculated. The average cell cycle profile is shown for individual samples, and experiments were performed in 3 biological replicates.

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: Histone signature in non-irradiated and irradiated cells without and with mutation or deletion in Suv39h1/h2 histone methyltransferases. Western blot analysis of γH2AX, H3K9me1, H3K9me2, H3K9me3, H3K9ac, H4K20me2, H4K20me3 and H4K20 acetylation. The levels of modified histones were normalized to that of total H3 histones. As DNA damage markers, 53BP1, MDC1 proteins, and the HP1β protein were studied and normalized to the level of α-tubulin. Protein levels were studied in ( A ) HAP1 wt and HAP1 cells with the mutation in the SUV39H1 gene and ( B ) in wt MEFs and Suv39h1/h2-deficient fibroblasts (MEFs). Non-irradiated cells and cells irradiated by 5 Gy of γ-rays (harvested 30 min and 24 hours after irradiation) were analyzed. ( C ) Cell cycle profiles were studied by flow cytometry in non-irradiated and γ-irradiated wt MEFs and non-irradiated and γ-irradiated Suv39h1/h2 dn mouse embryonic fibroblasts. Panel ( a ) shows a 30-min interval, and panel ( b ) shows a 24-h interval when the cells were harvested after irradiation (and related control samples). Using Mod-Fit software, the percentage of cells in G1 (red peak), S (dash blue peak) and G2-M (green peak) cell cycle phases was calculated. The average cell cycle profile is shown for individual samples, and experiments were performed in 3 biological replicates.

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Irradiation, Mutagenesis, Western Blot, Modification, Flow Cytometry, Cytometry, Software

    The nuclear distribution pattern of H4K20me3 (green) in UV-damaged chromatin . ( Aa ) Non-irradiated and γ-irradiated Suv39h1/h2 wt and ( Ab ) non-irradiated and γ-irradiated Suv39h1/h2 dn MEFs. DAPI (blue) was used as a counterstain. Scale bars represent 10 µm. ( B ) Quantification of H4K20me3 (green) shown in panels Aa, Ab. Quantification of H4K20me3 was performed according to the selected region of interests (ROIs, yellow lines). LAS AX software was used for analysis of fluorescence intensities. ( C ) Nuclear distribution pattern of H4K20me3 in ( a ) non-irradiated and ( c ) γ-irradiated Suv39h1/h2 wt and ( b ) non-irradiated and ( d ) γ-irradiated Suv39h1/h2 dn fibroblasts. ( D ) Quantification of fluorescence intensity of H4K20me3 is shown in panel Ca-d. Analysis by LAS AF software was performed in ( a ) non-irradiated and ( c ) γ-irradiated Suv39h1/h2 wt and ( b ) non-irradiated and ( d ) γ-irradiated Suv39h1/h2 dn MEFs. ( E ) Nuclear distribution of H4K20me3 (green) and the 53BP1 protein (red) in ( a ) non-irradiated and ( c ) γ-irradiated Suv39h1/h2 wt and ( b ) non-irradiated and ( d ) γ-irradiated Suv39h1/h2 dn cells.

    Journal: Aging (Albany NY)

    Article Title: H3K9me3 and H4K20me3 represent the epigenetic landscape for 53BP1 binding to DNA lesions

    doi: 10.18632/aging.101572

    Figure Lengend Snippet: The nuclear distribution pattern of H4K20me3 (green) in UV-damaged chromatin . ( Aa ) Non-irradiated and γ-irradiated Suv39h1/h2 wt and ( Ab ) non-irradiated and γ-irradiated Suv39h1/h2 dn MEFs. DAPI (blue) was used as a counterstain. Scale bars represent 10 µm. ( B ) Quantification of H4K20me3 (green) shown in panels Aa, Ab. Quantification of H4K20me3 was performed according to the selected region of interests (ROIs, yellow lines). LAS AX software was used for analysis of fluorescence intensities. ( C ) Nuclear distribution pattern of H4K20me3 in ( a ) non-irradiated and ( c ) γ-irradiated Suv39h1/h2 wt and ( b ) non-irradiated and ( d ) γ-irradiated Suv39h1/h2 dn fibroblasts. ( D ) Quantification of fluorescence intensity of H4K20me3 is shown in panel Ca-d. Analysis by LAS AF software was performed in ( a ) non-irradiated and ( c ) γ-irradiated Suv39h1/h2 wt and ( b ) non-irradiated and ( d ) γ-irradiated Suv39h1/h2 dn MEFs. ( E ) Nuclear distribution of H4K20me3 (green) and the 53BP1 protein (red) in ( a ) non-irradiated and ( c ) γ-irradiated Suv39h1/h2 wt and ( b ) non-irradiated and ( d ) γ-irradiated Suv39h1/h2 dn cells.

    Article Snippet: This procedure was modified for the antibody against histone H4K20me3 (#A-4048-050, Epigentek, Lab Mark a.s.).

    Techniques: Irradiation, Software, Fluorescence