gst tagged hdac proteins (Worthington Biochemical)

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Name:

Nuclease Micrococcal

Description:

Chromatographically purified to be essentially homogeneous chromatographically and electrophoretically SDS PAGE A lyophilized powder

Catalog Number:

ls004796

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Source:

Staphylococcus aureus (Strain Foggi)

Cas Number:

9013.53.0

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Structured Review

Worthington Biochemical gst tagged hdac proteins

AtTRB2 also interacts with HDA6, a RPD3-type histone deacetylase. ( A ) Western blot analysis showing the different patterns of interaction between AtTRB proteins and various <t>HDAC</t> proteins. PAGE shows the purified HDAC proteins used in this assay (first panel). ( B ) Scheme of various HDA6 truncated proteins fused to <t>GST.</t> HDA6 protein comprises three predicted regions: a conserved HDAC domain in the N-terminal region, a glutamate-rich region in the center of the protein and an aspartate-rich region in the C-terminal region. ( C ) Pull-down assay to map the regions of HDA6 that interact with AtTRB2. PAGE shows the purified HDA6-truncated proteins shown in (B). ( D ) Schematic map indicating the region of HDA6 involved in binding to AtTRB2. ( E ) Western blots demonstrating the interaction between AtTRB2 and HDA6 in vivo . Empty control and IP with IgG were used as negative controls. ( F ) HDA6-associated telomeric repeat signal was detected in an AtTRB2-dependent manner. ( G ) Quantification of multiple independent T-ChIP experiments as shown in (F). T-ChiP analysis was performed and values were calculated as described in the legend to Figure 4 .

Chromatographically purified to be essentially homogeneous chromatographically and electrophoretically SDS PAGE A lyophilized powder
https://www.bioz.com/result/gst tagged hdac proteins/product/Worthington Biochemical
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gst tagged hdac proteins - by Bioz Stars, 2021-01

88/100 stars

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Images

1) Product Images from "Telomere-binding protein regulates the chromosome ends through the interaction with histone deacetylases in Arabidopsis thaliana"

Article Title: Telomere-binding protein regulates the chromosome ends through the interaction with histone deacetylases in Arabidopsis thaliana

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkw067

Figure Legend Snippet: AtTRB2 also interacts with HDA6, a RPD3-type histone deacetylase. ( A ) Western blot analysis showing the different patterns of interaction between AtTRB proteins and various HDAC proteins. PAGE shows the purified HDAC proteins used in this assay (first panel). ( B ) Scheme of various HDA6 truncated proteins fused to GST. HDA6 protein comprises three predicted regions: a conserved HDAC domain in the N-terminal region, a glutamate-rich region in the center of the protein and an aspartate-rich region in the C-terminal region. ( C ) Pull-down assay to map the regions of HDA6 that interact with AtTRB2. PAGE shows the purified HDA6-truncated proteins shown in (B). ( D ) Schematic map indicating the region of HDA6 involved in binding to AtTRB2. ( E ) Western blots demonstrating the interaction between AtTRB2 and HDA6 in vivo . Empty control and IP with IgG were used as negative controls. ( F ) HDA6-associated telomeric repeat signal was detected in an AtTRB2-dependent manner. ( G ) Quantification of multiple independent T-ChIP experiments as shown in (F). T-ChiP analysis was performed and values were calculated as described in the legend to Figure 4 .

Techniques Used: Histone Deacetylase Assay, Western Blot, Polyacrylamide Gel Electrophoresis, Purification, Pull Down Assay, Binding Assay, In Vivo, Chromatin Immunoprecipitation

2) Product Images from "The Use of Mononucleosome Immunoprecipitation for Analysis of Combinatorial Histone Post-translational Modifications and Purification of Nucleosome-Interacting Proteins"

Article Title: The Use of Mononucleosome Immunoprecipitation for Analysis of Combinatorial Histone Post-translational Modifications and Purification of Nucleosome-Interacting Proteins

Journal: Frontiers in Cell and Developmental Biology

doi: 10.3389/fcell.2020.00331

Visualization of nuclei at different steps of the protocol using a phase-contrast microscope. Isolated nuclei at Step 6 (A) MNase-digested nuclei at Step 9 (B) TE-resuspended nuclei at Step 11 (C) .

Figure Legend Snippet: Visualization of nuclei at different steps of the protocol using a phase-contrast microscope. Isolated nuclei at Step 6 (A) MNase-digested nuclei at Step 9 (B) TE-resuspended nuclei at Step 11 (C) .

Techniques Used: Microscopy, Isolation

Isolation:

Article Title: Mcm1 regulates donor preference controlled by the recombination enhancer in Saccharomyces mating-type switching
Article Snippet: .. Nuclei were isolated, digested with micrococcal nuclease or DNaseI (Worthington), and DNA was purified as described ( ; ) with modifications detailed in . .. Naked DNA controls were obtained by digesting a PCR product.

Purification:

Concentration Assay:

Article Title: Elucidating combinatorial histone modifications and crosstalks by coupling histone-modifying enzyme with biotin ligase activity
Article Snippet: .. Nuclei were then resuspended in MNase Cutting Buffer containing 3 mM CaCl2 and 2 mM p -chloromercuri-phenylsulfonic acid (pCMPS, Toronto Research Labs; as phosphatase inhibitor), and digested with micrococcal nuclease (MNase, Worthington) at 37°C for 30 min at a concentration of 10U/107 cells. .. Digestion of chromatin by MNase was stopped by addition of EGTA to a final concentration of 5 mM, and equal volume of 2 x Lysis Buffer (30 mM HEPES, pH 7.9, 220 mM KCl, 3 mM MgCl2 , 2 mM EDTA, 0.4% Triton X-100, 20% glycerol) was added to the digested chromatin.

Incubation:

Article Title: Ribosomal Pausing at a Frameshifter RNA Pseudoknot Is Sensitive to Reading Phase but Shows Little Correlation with Frameshift Efficiency
Article Snippet: .. Unprotected mRNA was degraded by incubation with micrococcal nuclease (1 or 2 U/μl; Worthington) and RNase V1 (0.02 U/μl; Pharmacia) at 26°C for 30 min in the presence of 3.5 mM Mg(OAc)2 and 3 mM CaCl2 in a final reaction volume of 40 μl. .. Following addition of 60 μl of buffer T (20 mM HEPES, 150 mM KOAc, 10 mM Mg(OAc)2 , 5 mM EGTA, 2 mM dithiothreitol), the reaction mixture was overlayered onto a 60-μl cushion of 0.25 M sucrose in buffer T and subsequently centrifuged at 30 lb/in2 for 30 min in an A-110 rotor in a Beckman airfuge.

Article Title: A Key Role for Chd1 in Histone H3 Dynamics at the 3? Ends of Long Genes in Yeast
Article Snippet: .. 25–40 units (depending on yeast strain and cell density) of micrococcal nuclease (Worthington Biochemical) were added and spheroplasts were incubated at 37°C for 20 minutes. .. The digestion was halted by shifting the reactions to 4°C and adding 0.5 M EDTA to a final concentration of 10 mM.

Article Title: Development and Application of High-Content Biological Screening for Modulators of NET Production
Article Snippet: .. Neutrophils were pre-incubated at 37°C in 5% CO2 for 30 min prior to stimulation and further incubation at 37°C in 5% CO2 for up to 4 h. Post-incubation samples were treated for 10 min with 1 unit/ml micrococcal nuclease (MNase; Worthington 4789), centrifuged for 10 min at 1,800 rcf and 150 µl of supernatant was transferred to a black 96-well plate (Costar 3915). .. NET-DNA was stained with 1 µM Sytox green® (Invitrogen S7020) and quantified with a fluorometer (Berthold Twinkle2 LB970; ex:485 nm/em:525 nm).

Western Blot:

Article Title: Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes
Article Snippet: .. 6x gel loading dye (New England Biolabs, catalog number: B7042) Agarose (Fisher Scientific, catalog number: BP160) Ethidium bromide (Sigma-Aldrich, catalog number: E7637) CL2B Sepharose beads (Sigma-Aldrich, catalog number: CL2B300) Streptavidin Sepharose beads (GE Healthcare, catalog number: 17-5113-01) Insulin (Sigma-Aldrich, catalog number: I1882) Clarity Western ECL Substrate (Bio-Rad Laboratories, catalog number: 1705060) Primary antibody: anti-V5 tag (Thermo Fisher Scientific, Invitrogen, catalog number: R960-25) Secondary antibody: anti-Mouse IgG (Thermo Fisher Scientific, Invitrogen, catalog number: 31430) Nonfat dry milk (Walmart, Great Value) Yeast nitrogen base without amino acids (United States Biological, catalog number: Y2025) Glucose (Merck, Millipore Sigma, catalog number: DX0145) Micrococcal Nuclease (MNase) (Worthington Biochemical, catalog number: LS004797) Tris hydroxymethyl aminomethane (Tris) (Fisher Scientific, catalog number: BP152) Sodium tetraborate decahydrate (Na borate) (Sigma-Aldrich, catalog number: S9640) Sodium chloride (NaCl) (Fisher Scientific, catalog number: S271) Hydrochloric acid (HCl) (Fisher Scientific, catalog number: A144) Phenylmethylsulfonyl fluoride (PMSF) (RPI, catalog number: P20270) Sodium dodecylsulfate (SDS) (RPI, catalog number: L22010) Glycerol (Fisher Scientific, catalog number: G33) Bromophenol blue (Sigma-Aldrich, catalog number: B7021) 2-Mercaptoethanol (Sigma-Aldrich, catalog number: M3148) Tween 20 (Sigma-Aldrich, catalog number: P2287) Acetic acid, Glacial (Fisher Scientific, catalog number: A38) 40% acrylamide solution (Bio-Rad Laboratories, catalog number: 1610140) 2% Bis solution (Bio-Rad Laboratories, catalog number: 1610142) Ammonium persulfate (Fisher Scientific, catalog number: BP179) Tetramethylethylenediamine (TEMED) (Bio-Rad Laboratories, catalog number: 1610800) Sodium bicarbonate (NaHCO3 ) (Fisher Scientific, catalog number: S233) Sodium carbonate (Na2 CO3 ) (Fisher Scientific, catalog number: S263) Sodium hydroxide (NaOH) (Fisher Scientific, catalog number: S318) Methanol (Fisher Scientific, catalog number: A412) Synthetic media (see Recipes) MNase (see Recipes) Extraction (E) buffer (see Recipes) 2x SDS-sample buffer (SB) (see Recipes) Wash (W) buffer (see Recipes) 17% SDS-PAGE gel (see Recipes) 5% stacking gel (see Recipes) 1x SDS-running buffer (see Recipes) 40x Na carbonate buffer (see Recipes) Blotting buffer (see Recipes) TBST ( T ris- b uffered s aline + T ween 20) (see Recipes) .. Equipment P20, P200 and P1000 Pipettes (Gilson) Labquake Tube Shaker/Rotators (Thermo Fisher Scientific) Swing bucket rotor (Beckman Coulter, model: GH 3.8) Shaker incubator (INFORS HT) Mini-Beadbeater-96 (Bio Spec Product) Tabletop centrifuge (Beckman Coulter, model: Allegra 6R) Centrifuge for microcentrifuge tubes (Eppendorf, model: 5415 D) ChemiDoc Touch Imaging System (Bio-Rad Laboratories, model: ChemiDoc Touch Imaging System) Vortex-Genie 2 (Scientific Industries, model: Vortex-Genie 2) Water bath (Precision Scientific, catalog number: 66551) Vertical Mini-Gel systems (C.B.S.