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kinetic hdac2 assay kit  (BPS Bioscience)
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BPS Bioscience kinetic hdac2 assay kit
A) iPOND followed by analysis with mass spectrometry reveals decreased <t>HDAC2</t> in siPACS-1 cells. B) Level of class I HDACs in control and PACS-1 downregulated cells. C) iPOND assay reveals PACS-1 localization to the sites of DNA replication and it is important for HDAC2 and HDAC3 stability in the replication forks and to maintain chromatin regulation. D) Acetylation status of the HDAC2 and HDAC3 specific histone modification in control and PACS-1 down regulated cells. E) Depletion of PACS-1 leads to decreased HP1, marker for heterochromatin. F) MNase assay results of nuclei isolated from control vs PACS-1 knockdown cells. **p < 0.01.
Kinetic Hdac2 Assay Kit, supplied by BPS Bioscience, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A) iPOND followed by analysis with mass spectrometry reveals decreased HDAC2 in siPACS-1 cells. B) Level of class I HDACs in control and PACS-1 downregulated cells. C) iPOND assay reveals PACS-1 localization to the sites of DNA replication and it is important for HDAC2 and HDAC3 stability in the replication forks and to maintain chromatin regulation. D) Acetylation status of the HDAC2 and HDAC3 specific histone modification in control and PACS-1 down regulated cells. E) Depletion of PACS-1 leads to decreased HP1, marker for heterochromatin. F) MNase assay results of nuclei isolated from control vs PACS-1 knockdown cells. **p < 0.01.

Journal: Oncogene

Article Title: The Multifunctional Protein PACS-1 is Required for HDAC2 and HDAC3 Dependent Chromatin Maturation and Genomic Stability

doi: 10.1038/s41388-020-1167-x

Figure Lengend Snippet: A) iPOND followed by analysis with mass spectrometry reveals decreased HDAC2 in siPACS-1 cells. B) Level of class I HDACs in control and PACS-1 downregulated cells. C) iPOND assay reveals PACS-1 localization to the sites of DNA replication and it is important for HDAC2 and HDAC3 stability in the replication forks and to maintain chromatin regulation. D) Acetylation status of the HDAC2 and HDAC3 specific histone modification in control and PACS-1 down regulated cells. E) Depletion of PACS-1 leads to decreased HP1, marker for heterochromatin. F) MNase assay results of nuclei isolated from control vs PACS-1 knockdown cells. **p < 0.01.

Article Snippet: HDAC activity was measured with the fluorometric HDAC3 Activity Assay kit (EP1004, Sigma, St Louis, MO) and kinetic HDAC2 assay kit (53002, BPS biosciences, USA) according to the manufacturer’s instructions.

Techniques: Mass Spectrometry, Modification, Marker, Isolation

A) Cycloheximide treatment and the level of PACS-1, HDAC2 and HDAC3 in various time points. B) MG-132 treatment and the level of PACS-1, HDAC2 and HDAC3 in various time points. C) Co-IP studies of HeLa cell lysates using PACS-1 antibody showed PACS-1 binds to HDAC2 and HDAC3 and similarly (D) HDAC2 and HDAC3 antibodies pull down PACS-1.

Journal: Oncogene

Article Title: The Multifunctional Protein PACS-1 is Required for HDAC2 and HDAC3 Dependent Chromatin Maturation and Genomic Stability

doi: 10.1038/s41388-020-1167-x

Figure Lengend Snippet: A) Cycloheximide treatment and the level of PACS-1, HDAC2 and HDAC3 in various time points. B) MG-132 treatment and the level of PACS-1, HDAC2 and HDAC3 in various time points. C) Co-IP studies of HeLa cell lysates using PACS-1 antibody showed PACS-1 binds to HDAC2 and HDAC3 and similarly (D) HDAC2 and HDAC3 antibodies pull down PACS-1.

Article Snippet: HDAC activity was measured with the fluorometric HDAC3 Activity Assay kit (EP1004, Sigma, St Louis, MO) and kinetic HDAC2 assay kit (53002, BPS biosciences, USA) according to the manufacturer’s instructions.

Techniques: Co-Immunoprecipitation Assay

A) Full length (963a.a.) PACS-1 protein is divided in to N-term (1–499a.a.) and C-term (500–963a.a.). N-term is further divided into ARR (1–117a.a.), FBR (117–263a.a.) and MR (240–499a.a.). B) PACS-1 downregulated HeLa cells were complemented with Myc-tagged N-terminal, C-terminal and full length PACS-1. Interestingly, full length PACS-1-Myc and N-terminal PACS-1-Myc brought back the HDAC2 and HDAC3 levels and reduced the γH2AX formation in PACS-1 downregulated cells. C) IP results show that FBR region of PACS-1-Myc binds to the HDAC3.

Journal: Oncogene

Article Title: The Multifunctional Protein PACS-1 is Required for HDAC2 and HDAC3 Dependent Chromatin Maturation and Genomic Stability

doi: 10.1038/s41388-020-1167-x

Figure Lengend Snippet: A) Full length (963a.a.) PACS-1 protein is divided in to N-term (1–499a.a.) and C-term (500–963a.a.). N-term is further divided into ARR (1–117a.a.), FBR (117–263a.a.) and MR (240–499a.a.). B) PACS-1 downregulated HeLa cells were complemented with Myc-tagged N-terminal, C-terminal and full length PACS-1. Interestingly, full length PACS-1-Myc and N-terminal PACS-1-Myc brought back the HDAC2 and HDAC3 levels and reduced the γH2AX formation in PACS-1 downregulated cells. C) IP results show that FBR region of PACS-1-Myc binds to the HDAC3.

Article Snippet: HDAC activity was measured with the fluorometric HDAC3 Activity Assay kit (EP1004, Sigma, St Louis, MO) and kinetic HDAC2 assay kit (53002, BPS biosciences, USA) according to the manufacturer’s instructions.

Techniques:

A and B) ADmut, GGAmut and S278A of GFP-PACS-1 are able to complement HDAC2 and HDAC3 expression. C) IDmut of GFP-PACS-1 is able to complement HDAC2 and HDAC3 expression. D and E) HDACmut of GFP-PACS-1 is unable to complement HDAC2 and HDAC3 expression. F) Affinity purified HDAC was incubated with GST or GST-PACS-1 FBR or GST-PACS-1 FBR HDACmut. GST proteins were captured and bound HDAC3 was detected by western blot. **p < 0.01.

Journal: Oncogene

Article Title: The Multifunctional Protein PACS-1 is Required for HDAC2 and HDAC3 Dependent Chromatin Maturation and Genomic Stability

doi: 10.1038/s41388-020-1167-x

Figure Lengend Snippet: A and B) ADmut, GGAmut and S278A of GFP-PACS-1 are able to complement HDAC2 and HDAC3 expression. C) IDmut of GFP-PACS-1 is able to complement HDAC2 and HDAC3 expression. D and E) HDACmut of GFP-PACS-1 is unable to complement HDAC2 and HDAC3 expression. F) Affinity purified HDAC was incubated with GST or GST-PACS-1 FBR or GST-PACS-1 FBR HDACmut. GST proteins were captured and bound HDAC3 was detected by western blot. **p < 0.01.

Article Snippet: HDAC activity was measured with the fluorometric HDAC3 Activity Assay kit (EP1004, Sigma, St Louis, MO) and kinetic HDAC2 assay kit (53002, BPS biosciences, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Affinity Purification, Incubation, Western Blot

A) PACS-1 downregulated HeLa cells were complemented with GFP-PACS-1 HDACmut. B) Fiber Assay and C) Chromosomal Aberrations in PACS-1 downregulated HeLa cells complemented with GFP-PACS-1 HDACmut. D) A hypothetical model showing PACS-1 protein, which is mostly present in the cytoplasm, accumulates into the nucleus during S-phase of the cell cycle and during genotoxic stress. Nuclear localized PACS-1 then interacts and stabilizes HDAC2 and HDAC3, which otherwise induces replication stress and genomic instability. *p < 0.05; ***p < 0.001; **** p < 0.0001.

Journal: Oncogene

Article Title: The Multifunctional Protein PACS-1 is Required for HDAC2 and HDAC3 Dependent Chromatin Maturation and Genomic Stability

doi: 10.1038/s41388-020-1167-x

Figure Lengend Snippet: A) PACS-1 downregulated HeLa cells were complemented with GFP-PACS-1 HDACmut. B) Fiber Assay and C) Chromosomal Aberrations in PACS-1 downregulated HeLa cells complemented with GFP-PACS-1 HDACmut. D) A hypothetical model showing PACS-1 protein, which is mostly present in the cytoplasm, accumulates into the nucleus during S-phase of the cell cycle and during genotoxic stress. Nuclear localized PACS-1 then interacts and stabilizes HDAC2 and HDAC3, which otherwise induces replication stress and genomic instability. *p < 0.05; ***p < 0.001; **** p < 0.0001.

Article Snippet: HDAC activity was measured with the fluorometric HDAC3 Activity Assay kit (EP1004, Sigma, St Louis, MO) and kinetic HDAC2 assay kit (53002, BPS biosciences, USA) according to the manufacturer’s instructions.

Techniques: