DOCSLIB.ORG

Chromatin Immunoprecipitation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Chromatin Immunoprecipitation CHROMATIN IMMUNOPRECIPITATION Diagenode provides complete solutions for ChIP, facilitating workflows and ensuring consistent results in every experiment PAGE 2 DIAGENODE CHROMATIN IMMUNOPRECIPITATION Complete solution for your ChIP Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has everything you need to make ChIP easy and convenient while ensuring consistent data between samples and experiments. As an expert in the field of epigenetics, Diagenode is committed to providing complete solutions from chromatin shearing reagents, shearing instruments such as the Bioruptor® (the gold standard for chromatin shearing), ChIP kits, the largest number of validated and trusted antibodies on the market, and the SX-8G IP-Star® Compact Automated System to achieve unparalleled productivity and reproducibility. INDUSTRY-LEADING CHIP KITS Diagenode offers a broad portfolio of extensively validated, high-quality ChIP kits. Choose the one that fits with your specific experimental needs (page 8). CHIP-SEQ AND CHIP-GRADE ANTIBODIES Diagenode focuses exclusively on epigenetics, providing the largest selection of industry-leading validated antibodies with dedicated technical expertise and support. Our antibodies are the only ones on the market validated in-house for ChIP-seq. Additonally, Diagenode offers premium antibodies which have reached the highest level of validation from extensive in-house validation in combination with numerous collaborations with the EU community of epigenetic experts (page 22). BIORUPTOR® SONICATION DEVICE The Bioruptor is now the industry’s standard and most cited tool for DNA shearing, chromatin shearing and cell lysis*, and is validated for Next-Generation Sequencing library preparation, ChIP, and DNA methylation studies (page 30). AUTOMATED SOLUTIONS The SX-8G IP-Star® Compact Automated System is a simple yet robust automated bench-top instrument that standardizes different epigenetic applications (e.g. ChIP), delivers consistent data and increases your lab’s productivity (page 34). * See list of selected publications on page 31 Innovating Epigenetic Solutions PAGE 3 About Epigenetics Table of contents In 2003, U.S. National Human Genome Research Institute About Epigenetics (NHGRI) director Francis Collins declared, “The HGP has Epigenetic Modifications of Histones . 4 been an amazing adventure into ourselves, to understand our own DNA instruction book, the shared inheritance of all What is ChIP? . 6 humankind.” Raw sequences of an estimated 30,000 genes Diagenode’s ChIP-seq workflow . 7 coding for various cellular functions were identified. STEP 1: Chromatin Shearing Optimization Since that time, much of the focus in human molecular Chromatin Shearing Optimization kits . 8 biology has been on the study of gene regulation in response to intra- and extra-cellular stimuli. This led to one of the STEP 2: Sonication challenges in modern biology: understanding the processes Overview . 9 and mechanisms that allow a single cell to become a ® complete organism composed of hundreds of individual cell Bioruptor Gold standard for chromatin shearing . 10 types, all of which still contain the same DNA. Researchers Efficient chromatin shearing with the Bioruptor® Plus . 11 soon discovered that chromatin could be likened to an The shearing device of choice . 12 “instruction book” which needs to be opened first so that Bioruptor® Models . 13 the words (i.e. the genes), can be read and interpreted. The epigenetic regulation of gene expression refers to STEP 3: Immunoprecipitation this phenomenon. The term epigenetics defines what is Antibodies . 14 happening on the chromatin, the physical support of genes. Manual ChIP kits . 32 Epigenetics is also defined as the study of heritable changes Automated ChIP kits . 37 in phenotype that are independent of changes in the DNA Automation . 40 sequence. “Epigenetic landscape” was a definition introduced by Conrad H. Waddington in the first part of the 20th century STEP 4: DNA Purification as “... the interactions of genes with their environment which IPure kit . 44 bring the phenotype into being” to describe the phenomena connecting genotype to phenotype and describing the STEP 5: Library Preparation process of differentiation. MicroPlex Library Preparation kit . 45 iDeal Library Preparation kit . 46 About Diagenode . 47 Diagenode Headquarters . Back Cover © 2016 Diagenode SA and Diagenode Inc. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, stored in retrieval systems, or translated into any language or computer language, in any form or by any means: electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without prior written permission from Diagenode SA (hereinafter, “Diagenode”) . The information in this guide is subject to change without notice. Diagenode and/or its affiliates reserve the right to change products and services at any time to incorporate the latest technological developments. Although this guide has been prepared with every precaution to ensure accuracy, Diagenode and/or its affiliates assume no liability for any errors or omissions, nor for any damages resulting from the application or use of this information. Diagenode welcomes customer input on corrections and suggestions for improvement. The trademarks mentioned herein are the property of Diagenode or their respective owners. Bioruptor® and IP-Star® are registered trademarks of Diagenode SA. Illumina is a registered trademark of Illumina Inc. Ion Torrent and Personal Genome Machine are trademarks of Life Technologies Corporation. www.diagenode.com | PAGE 4 DIAGENODE CHROMATIN IMMUNOPRECIPITATION In 1942, Waddington stated “... it is possible that an adaptive Three major levels of epigenetic changes are described response can be fixed without waiting for the occurrence today: of a mutation...” explaining the plasticity of epigenetic processes, and thereby framing their potential heritability, Chemical modifications at the level of the or long-term stability. nucleotides, which include DNA methylation and RNA interference Epigenetic machinery gives an organism with the molecular Modifications at the level of histones that include resources a quick response to environmental conditions post-translational change of histone proteins and with a steady modification in gene expression. the incorporation of histone variants Visual representation of the epigenetic landscape. ATP-dependent processes that regulate the access to nucleosomal DNA (nucleosome remodelling) Epigenetic mechanisms are made of a highly complex and interlacing network, incorporating all three levels of possible epigenetic modifications. These mechanisms include multiple positive and negative feed forward and feedback pathways that span both the DNA and the histone level and the higher order of chromatin structure. A number of epigenetic modifications are found at both of these levels due to mechanisms which still remain poorly understood. Epigenetic Modifications of Histones Post-translational histone modifications affinity between the histone tail and the negatively charged DNA. Histones can become acetylated on lysine (K) (PTMs) residues. Histone acetylation are regulated by the histone acetyltransferases (HATs) and two functional categories of Due to their chemical properties, PTMs modify the histone deacetylases (HDACs): 1) eleven zinc-dependent condensation of chromatin and therefore the accessibility HDACs (class I, IIa, IIb, and IV HDACs) and 2) the sirtuins of DNA to transcriptional machinery. The majority of histone (seven class III NAD -dependent HDACs). Functional impact PTMs arises on the NH2-terminal tail, although on the of some acetylateable residues is more substantial than COOH-terminal tail of the core histones. PTMs consist of others. For example, in yeast (S. cerevisae), H4K16 regulates acetylation, methylation, phosphorylation, ubiquitination, its own subset of genes, while other genes are co-regulated and sumoylation. PTMs can occur on all histones. by all of H4K12, K8, and K5, but not by K16. Histones are basic proteins that regulate the compaction of the chromatin. Histones consist of a globular histone 2. Methylation core and a NH2-terminal histone tail, protruding out of the rigid histone-DNA assemblage, called the nucleosome. This epigenetic modification has been shown to associate A nucleosome, the 10 nm thick primary structure of the with both transcribed and silenced genes. Histones can be chromatin is made of an octamer of the four core histones methylated on both lysine and arginine (R) residues. Lysine H2A, H2B, H3, and H4 in duplicates, around which 147 residues can be mono-, di-, and trimethylated and arginine base pairs of DNA are wrapped. Nucleosomes are bonded residues can be mono- and dimethylated. Effector proteins, together by the linker histone H1, which is not part of the such as transcriptional co-activators, recognize mono-, di-, nucleosome as such. and trimethylated epitopes with different affinities. Thus it is most likely that the various degrees of histone methylation influence gene expression regulation (e.g. trimethylated 1. Acetylation H3K4 is exclusively associated with expressed genes Histone acetylation has been correlated to transcriptional whereas monomethylated H3K4 has been associated with activation. Acetyl group addition neutralizes the positive both expressed and repressed genes). charge of the -amino group of the lysine, which decreases Innovating Epigenetic Solutions PAGE 5 4. Ubiquitination
Load more

Recommended publications
  • Epigenetic Regulation of Muscle Stem Cell Expansion
    Epigenetic regulation of muscle stem cell expansion INAUGURAL-DISSERTATION zur Erlangung des Doktorgrades der Naturwissenschaften - Doctor rerum naturalium - (Dr. rer. nat.) vorgelegt dem Fachbereich für Biologie und Chemie (FB 08) der Justus-Liebig-Universität Gießen eingereicht von Ting Zhang Gießen, 2015 Die vorliegende Arbeit wurde am Max-Planck-Institut für Herz- und Lungenforschung, W.G. Kerckhoff-Institut in Bad Nauheim angefertigt. Erstgutachter: Prof. Dr. Dr. Thomas Braun Abteilung Entwicklung und Umbau des Herzens Max-Planck-Institut für Herz- und Lungenforschung Ludwigstraße 43, 61231 Bad Nauheim Zweitgutachter: Prof. Dr. Lienhard Schmitz Biochemisches Institut Fachbereich Medizin Justus-Liebig-Universität Gießen Friedrichstrasse 24, 35392 Giessen Disputation am 10.Juni 2015 EIDESSTATTLICHE ERKLÄRUNG „Ich erkläre: Ich habe die vorgelegte Dissertation selbständig und ohne unerlaubte fremde Hilfe und nur mit den Hilfen angefertigt, die ich in der Dissertation angegeben habe. Alle Textstellen, die wörtlich oder sinngemäß aus veröffentlichten Schriften entnommen sind, und alle Angaben, die auf mündlichen Auskünften beruhen, sind als solche kenntlich gemacht. Bei den von mir durchgeführten und in der Dissertation erwähnten Untersuchungen habe ich die Grundsätze guter wissenschaftlicher Praxis, wie sie in der „Satzung der Justus-Liebig-Universität Gießen zur Sicherung guter wissenschaftlicher Praxis“ niedergelegt sind, eingehalten.“ Bad Nauheim, den ZUSAMMENFASSUNG ZUSAMMENFASSUNG Stammzellen der adulten Skelettmuskulatur, auch als
    [Show full text]
  • Subcloning, Expression, and Enzymatic Study of PRMT5
    Georgia State University ScholarWorks @ Georgia State University Biology Theses Department of Biology Summer 7-12-2010 Subcloning, Expression, and Enzymatic Study of PRMT5 Ran Guo Georgia State University Follow this and additional works at: https://scholarworks.gsu.edu/biology_theses Part of the Biology Commons Recommended Citation Guo, Ran, "Subcloning, Expression, and Enzymatic Study of PRMT5." Thesis, Georgia State University, 2010. https://scholarworks.gsu.edu/biology_theses/26 This Thesis is brought to you for free and open access by the Department of Biology at ScholarWorks @ Georgia State University. It has been accepted for inclusion in Biology Theses by an authorized administrator of ScholarWorks @ Georgia State University. For more information, please contact [email protected]. SUBCLONING, EXPRESSION, AND ENZYMATIC STUDY OF PRMT5 by RAN GUO Under the Direction of Yujun George Zheng ABSTRACT Protein arginine methyltransferases (PRMTs) mediate the transfer of methyl groups to arginine residues in histone and non-histone proteins. PRMT5 is an important member of PRMTs which symmetrically dimethylates arginine 8 in histone H3 (H3R8) and arginine 3 in histone H4 (H4R3). PRMT5 was reported to inhibit some tumor suppressors in leukemia and lymphoma cells and regulate p53 gene, through affecting the promoter of p53. Through methylation of H4R3, PRMT5 can recruit DNA-methyltransferase 3A (DNMT3A) which regulates gene transcription. All the above suggest that PRMT5 has an important function of suppressing cell apoptosis and is a potential anticancer target. Currently, the enzymatic activities of PRMT5 are not clearly understood. In our study, we improved the protein expression methodology and greatly enhanced the yield and quality of the recombinant PRMT5.
    [Show full text]
  • UNIVERSITY of CALIFORNIA, IRVINE Combinatorial Regulation By
    UNIVERSITY OF CALIFORNIA, IRVINE Combinatorial regulation by maternal transcription factors during activation of the endoderm gene regulatory network DISSERTATION submitted in partial satisfaction of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biological Sciences by Kitt D. Paraiso Dissertation Committee: Professor Ken W.Y. Cho, Chair Associate Professor Olivier Cinquin Professor Thomas Schilling 2018 Chapter 4 © 2017 Elsevier Ltd. © 2018 Kitt D. Paraiso DEDICATION To the incredibly intelligent and talented people, who in one way or another, helped complete this thesis. ii TABLE OF CONTENTS Page LIST OF FIGURES vii LIST OF TABLES ix LIST OF ABBREVIATIONS X ACKNOWLEDGEMENTS xi CURRICULUM VITAE xii ABSTRACT OF THE DISSERTATION xiv CHAPTER 1: Maternal transcription factors during early endoderm formation in 1 Xenopus Transcription factors co-regulate in a cell type-specific manner 2 Otx1 is expressed in a variety of cell lineages 4 Maternal otx1 in the endodermal conteXt 5 Establishment of enhancers by maternal transcription factors 9 Uncovering the endodermal gene regulatory network 12 Zygotic genome activation and temporal control of gene eXpression 14 The role of maternal transcription factors in early development 18 References 19 CHAPTER 2: Assembly of maternal transcription factors initiates the emergence 26 of tissue-specific zygotic cis-regulatory regions Introduction 28 Identification of maternal vegetally-localized transcription factors 31 Vegt and OtX1 combinatorially regulate the endodermal 33 transcriptome iii
    [Show full text]
  • Reversible Histone Glycation Is Associated with Disease-Related Changes in Chromatin Architecture
    ARTICLE https://doi.org/10.1038/s41467-019-09192-z OPEN Reversible histone glycation is associated with disease-related changes in chromatin architecture Qingfei Zheng 1, Nathaniel D. Omans1,2, Rachel Leicher3,4, Adewola Osunsade 1,4, Albert S. Agustinus 1, Efrat Finkin-Groner1, Hannah D’Ambrosio1, Bo Liu5, Sarat Chandarlapaty 5, Shixin Liu3 & Yael David1,4,6,7 Cellular proteins continuously undergo non-enzymatic covalent modifications (NECMs) that accumulate under normal physiological conditions and are stimulated by changes in the 1234567890():,; cellular microenvironment. Glycation, the hallmark of diabetes, is a prevalent NECM asso- ciated with an array of pathologies. Histone proteins are particularly susceptible to NECMs due to their long half-lives and nucleophilic disordered tails that undergo extensive regulatory modifications; however, histone NECMs remain poorly understood. Here we perform a detailed analysis of histone glycation in vitro and in vivo and find it has global ramifications on histone enzymatic PTMs, the assembly and stability of nucleosomes, and chromatin archi- tecture. Importantly, we identify a physiologic regulation mechanism, the enzyme DJ-1, which functions as a potent histone deglycase. Finally, we detect intense histone glycation and DJ-1 overexpression in breast cancer tumors. Collectively, our results suggest an additional mechanism for cellular metabolic damage through epigenetic perturbation, with implications in pathogenesis. 1 Chemical Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. 2 Tri-Institutional Training Program in Computational Biology and Medicine, New York, NY 10065, USA. 3 Laboratory of Nanoscale Biophysics and Biochemistry, Rockefeller University, New York, NY 10065, USA. 4 Tri-institutional PhD Program in Chemical Biology, New York, NY 10065, USA.
    [Show full text]
  • PRMT5 Modulates the Metabolic Response to Fasting Signals
    PRMT5 modulates the metabolic response to fasting signals Wen-Wei Tsaia, Sherry Niessenb, Naomi Goebela, John R. Yates IIIb, Ernesto Guccionec,d, and Marc Montminya,1 aThe Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037; bDepartment of Chemical Physiology, The Scripps Research Institute, La Jolla, CA 92037; cInstitute of Molecular and Cell Biology, Proteos, Singapore 138673; dDepartment of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119074 Contributed by Marc Montminy, March 11, 2013 (sent for review February 20, 2013) Under fasting conditions, increases in circulating glucagon maintain sites. The results provide a mechanism to explain how latent cy- glucose balance by promoting hepatic gluconeogenesis. Triggering toplasmic regulators such as CRTC2 may contribute to signaling of the cAMP pathway stimulates gluconeogenic gene expression in the nucleus through their association with chromatin mod- through the PKA-mediated phosphorylation of the cAMP response ifying enzymes. element binding (CREB) protein and via the dephosphorylation of the latent cytoplasmic CREB regulated transcriptional coactivator 2 Results (CRTC2). CREB and CRTC2 activities are increased in insulin resis- In mass spectroscopy studies using epitope-tagged CRTC2 to tance, in which they promote hyperglycemia because of constitu- identify relevant interacting proteins, we recovered the protein tive induction of the gluconeogenic program. The extent to which arginine methyltransferase 5 (PRMT5) (Fig. S1 A and B). We CREB and CRTC2 are coordinately up-regulated in response to glu- confirmed the CRTC2:PRMT5 interaction in coimmunopreci- cagon, however, remains unclear. Here we show that, following its pitation studies with epitope-tagged proteins (Fig.
    [Show full text]
  • H3r8me2(Asym) Polyclonal Antibody - Classic
    H3R8me2(asym) polyclonal antibody - Classic Cat. No. C15410286 Specificity: Human, mouse, C. elegans, rat, chicken, Type: Polyclonal Xenopus, Drosophila, plant Source: Rabbit Purity: Affinity purified Lot #: 001 Storage: Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. Size: 50 µg Precautions: This product is for research use only. Not for Concentration: 0.84 µg/µl use in diagnostic or therapeutic procedures. Applications Suggested dilution Results ChIP 2-5 µg/million cells Figure 1 Immunohistochemistry 1:100 IF 1:200 Figure 2 Western blot 1:500 Figure 3 Target description Chromatin is the arrangement of DNA and proteins in which chromosomes are formed. Correspondingly, chromatin is formed from nucleosomes, which are comprised of a set of four histone proteins (H2A, H2B, H3, H4) wrapped with DNA. Chromatin is a very dynamic structure in which numerous post-translational modifications work together to activate or repress the availability of DNA to be copied, transcribed, or repaired. These marks decide which DNA will be open and commonly active (euchromatin) or tightly wound to prevent access and activation (heterochromatin). Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. In particular, dimethylation of H3 Arg8 (H3 R8Me2) is known as a mark of transcriptional repression. The protein arginine methyltransferases PRMT5 and PRMT2 can both methylate Arg8, with PRMT2 specifically methylating in an asymmetric manner. In addition, asymmetric dimethylation of Arg8 inhibits H3K9 methylation by G9a, but symmetric Arg8Me2 does not. 1 Results Figure 1.
    [Show full text]
  • Symmetric-Di-Methyl-Histone H3 (Arg8) Rabbit
    Symmetric -Di-methyl-Histone H3 (Arg8) rabbit pAb ( H3R8me2(sy) pAb) www.ptm-biolab.com.cn Cat#: PTM-672 [email protected] Species: Rabbit 0571-2883 3567 Size: 100 μl Form: supplied in liquid form Recommend Species Swissprot Molecular Application Immunogen dilution Reactivity ID Weight H3R8me2(sy) WB, Dot, ELISA 1:2000 for WB H, M, R, Mk P68431 17 KDa -KLH **Species reactivity is determined by WB. Kept at -20oC after reconstituted. *** Anti-rabbit secondary antibodies must be used to detect this antibody. Source/Purification: This product is produced by immunizing rabbits with a synthetic di-methyl(sy) peptide corresponding to residues surrounding Arg8 of human histone H3. Antibodies are purified by protein A-conjugated agarose followed by di-methylated(sy) histone H3 (Arg8) peptide affinity chromatography. Recommended Applications: ELISA, Dot blot, Western blot. Recommended antibody dilution: WB: 1:2000 NOTE: For WB, incubate membrane with diluted antibody in 5% nonfat milk, 1 x TBS, 0.1% Tween- 20 for two hours at room temperature with gentle shaking. Prepare working dilution immediately before use. Use at an assay dependent concentration. Optimal dilutions/concentrations should be determined by the end user. Not yet tested in other applications Scientific Description: Proteins post-translational Modification (PTM) is an important reversible event controlling protein activity. The amino-terminal tails of core histones undergo multiple modifications in multiple sites, termed as “histone code” or “epigenetic code”. Lysine methylation or arginine methylation in core histones is a major determinant for the formation of active and inactive regions of the genome and therefore plays vital roles in multiple cellular events.
    [Show full text]
  • H3r8me2(Asym) | Histone H3 (Asym-Dimethylated Arg8) Product Information
    Product no AS16 3177 H3R8me2(asym) | Histone H3 (asym-dimethylated Arg8) Product information Immunogen KLH-conjugated synthetic peptide Host Rabbit Clonality Polyclonal Purity Affinity purified serum Format Liquid Quantity 50 µg Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes. Additional information This antibody preparation is provided in 20 mM Potassium Phosphate pH 7.2, 150 mM NaCl, 0,01% sodium azide and 30% glycerol. Application information Recommended dilution 2 ug (ChIP), 1: 100 (IF), 1: 100 (IHC), 1:500 (WB) Expected | apparent 15 kDa MW Confirmed reactivity Caenorhabditis elegans, Human Predicted reactivity Chicken, Drosophila melanogaster, Mouse, Plant, Rat, Xenopus sp. Not reactive in No confirmed exceptions from predicted reactivity are currently known. application example Chromatin Immunoprecipitation using anti-H3R8me2(asym) antibodies. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 µg of H3R8me2(asym) and 20ul of magnetic beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR, and normalized to the input chromatin. Immunofluorescence using anti-H3R8me2(asym) antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: used at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10 000 for 45 min at RT. Localization: H3R8me2(asym) is nuclear and chromosomal. Staining: H3R8me2(asym) is expressed in green and the nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight® 594 (red).
    [Show full text]
  • Anti-Symmetric Di-Methyl-Histone H3 (Arg2) Mouse Mab
    Anti-Symmetric Di-Methyl-Histone H3 (Arg2) Mouse mAb Catalog # PTM-693 General Information Images Host species Mouse Dot Blot Clonality Monoclonal Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:1000 Clone number / Primary ab incubation condition: 2 hours at room temperature Synonym H3R2me2 (sy) Secondary ab: Goat Anti-Mouse IgG H&L Immunogen UniProt MW (kDa) Applications (HRP) species Immunogen peptide quantity: 1 ng, 4 ng, 16 ng Human P68431 17 WB Exposure time: 60 s The list of peptides is included in the table below: No. Peptide No. Peptide Product Usage Information Lane 1 H3R2me2 (sy) Lane 2 H3R2me1 Application Dilution Recommended species Lane 3 H3R2me2 (asy) Lane 4 H3K4ac Human, Mouse, Rat, Lane 5 H3K4cr Lane 6 H3K4hib WB 1:500-1:2000 Lane 7 H3K4me1 Lane 8 H3K4me2 Chlorocebus aethiops Lane 9 H3K4me3 Lane 10 H3R8me1 Lane 11 H3R8me2 Lane 12 H3R8me2 (asy) Properties Lane 13 H3R2un Storage Store at -20 °C. Avoid freeze / thaw cycles. WB Blocking buffer: 5% NFDM/TBST Stability Stable for 12 months from date of receipt / reconstitution Primary ab dilution: 1:2000 Constituents PBS, Glycerol, BSA Primary ab incubation condition: 2 hours at room temperature Purity Antibodies are purified by protein G-conjugated agarose Secondary ab: Goat Anti-Mouse IgG H&L followed by di-methylated(sy) histone H3 (Arg2) peptide (HRP) Lysate: MCF-7, BRL, COS-7 affinity chromatography. Protein loading quantity: 20 μg Isotype IgG Exposure time: 60 s Predicted MW: 17 kDa Target Information Observed MW: 17 kDa Function Histone post-translational modifications (PTMs) are key mechanisms of epigenetics that modulate chromatin Research Use structures, termed as “histone code”.
    [Show full text]
  • Epigenetic Control of Reprogramming and Cellular Differentiation
    Comparative and Functional Genomics EPIGENETIC CONTROL OF REPROGRAMMING AND CELLULAR DIFFERENTIATION Guest Editors: Lucia Latella, Daniela Palacios, Sonia Forcales, and Pier Lorenzo Puri Epigenetic Control of Reprogramming and Cellular Differentiation Comparative and Functional Genomics Epigenetic Control of Reprogramming and Cellular Differentiation Guest Editors: Lucia Latella, Daniela Palacios, Sonia Forcales, and Pier Lorenzo Puri Copyright © 2012 Hindawi Publishing Corporation. All rights reserved. This is a special issue published in “Comparative and Functional Genomics.” All articles are open access articles distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Editorial Board Allan Bradley, UK S. Hagemann, Austria E. Pasyukova, Russia Kevin Brindle, UK H. Heng, USA G. Pesole, Italy J. Camonis, France E. Hovig, Norway G. Piaggio, Italy S. Liang Chen, Taiwan Yeon-Su Lee, Korea E. M. Reis, Brazil P. V. Chou da r y, USA Peter Little, Australia Brian Wigdahl, USA M. A. Collart, Switzerland Ali Masoudi-Nejad, Iran W. Zhang, USA Ian Dunham, UK Giuliana Napolitano, Italy Jinfa Zhang, USA Soraya E. Gutierrez, Chile Ferenc Olasz, Hungary M. Hadzopoulou-Cladaras, Greece John Parkinson, Canada Contents Epigenetic Control of Reprogramming and Cellular Differentiation, Lucia Latella, Daniela Palacios, Sonia Forcales, and Pier Lorenzo Puri Volume 2012, Article ID 538639, 2 pages A Systems Approach and Skeletal Myogenesis,
    [Show full text]
  • Anti-H3r8me2(Asym) Antibody
    FOR RESEARCH USE ONLY! 01/20 Anti-H3R8me2(asym) Antibody CATALOG NO.: A2047-100 (100 µl) BACKGROUND DESCRIPTION: Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack poly A tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. H3.4; H3/g; H3FT; H3t; HIST3H3; Histone H3; HIST1H3A ALTERNATE NAMES: ANTIBODY TYPE: Polyclonal HOST/ISOTYPE: Rabbit / IgG IMMUNOGEN: A synthetic methylated peptide targeting residues around Arginine 8 of human histone H3 MOLECULAR WEIGHT: 18 kDa PURIFICATION: Affinity purified FORM: Liquid FORMULATION: In PBS with 0.02% sodium azide, 50% glycerol, pH7.3 SPECIES REACTIVITY: Human, Mouse, Rat STORAGE CONDITIONS: Store at -20ºC. Avoid freeze / thaw cycles APPLICATIONS AND USAGE: WB 1:500 - 1:2000, IF 1:50 - 1:200 Note: This information is only intended as a guide. The optimal dilutions must be determined by the user Western blot analysis of H3R8me2(asym) expression in Dot-blot analysis of methylation peptides using Anti- HeLa cells and H3 protein.
    [Show full text]
  • Theranostics CAPG Enhances Breast Cancer Metastasis by Competing
    Theranostics 2018, Vol. 8, Issue 9 2549 Ivyspring International Publisher Theranostics 2018; 8(9): 2549-2564. doi: 10.7150/thno.22523 Research Paper CAPG enhances breast cancer metastasis by competing with PRMT5 to modulate STC-1 transcription Sheng Huang1,2#, Yayun Chi1,#, Yi Qin3,#, Ziliang Wang4, Bingqiu Xiu1, Yonghui Su1, Rong Guo1, Liang Guo1, Hefen Sun1, Chujia Zeng1, Shuling Zhou5, Xin Hu1, Sheng Liu6, Zhimin Shao1, Zhaohui Wu7, Wei Jin1,, Jiong Wu1, Department of Breast Surgery, Breast Cancer Institute, Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, People's Republic of China The Second Department of Breast Surgery, The Third Affiliated Hospital of Kunming Medical University (Tumor Hospital of Yunnan Province), Kunming, Yunnan, People's Republic of China Department of Pancreas & Hepatobiliary Surgery, Pancreas & Hepatobiliary Cancer Institute, Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, People's Republic of China Cancer Institute, Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, People’s Republic of China Department of Pathology, Shanghai Cancer Center, Fudan University, Shanghai, People's Republic of China Department of Breast Surgery and Pharmacology Laboratory of Traditional Chinese Medicine, Long Hua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, People's Republic of China Department of Pathology and Laboratory Medicine; Center for Cancer Research, University of Tennessee Health Science Center, Memphis, Tennessee # These authors contributed equally to this work. Corresponding author: Jiong Wu, Department of Breast Surgery, Building 2, No. 270 Dong An Road, Shanghai, 200032 Email: [email protected]; Tel: 86-21-64175590-3423; Fax: 86-21-64172585 or Wei Jin, Breast Cancer Institute, Building 7, No.
    [Show full text]