17-408 EZ-Magna ChIP™ A - Chromatin Immunoprecipitation Kit

17-408
22 assays  Kit capacity: 22 chromatin immunoprecipitation assays
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      For target-specific spike-in controls that make ChIP experiments more quantitative and accurate, Click on the Related Product & Applications tab above.
      Description
      Catalogue Number17-408
      Trade Name
      • Magna ChIP
      DescriptionEZ-Magna ChIP™ A - Chromatin Immunoprecipitation Kit
      OverviewChromatin Immunoprecipitation (ChIP) is an important technique allowing the researcher to analyze in vivo interactions of proteins with genomic DNA. Any chromatin-associated or DNA binding protein can be analyzed with this technique, provided a good antibody to the protein exists. One can measure different proteins localized to a specific region of the genome, or the genome wide distribution of a specific protein. Another powerful application of this technique is to analyze changes in histone modifications that correlate with processes like transcription, mitosis or DNA repair.

      Features & Benefits:
      Faster: Magnetic protein A beads allow for the entire ChIP protocol to be done in as little as a day! All reagents to process your samples are included - you don't have to spend valuable time making them.
      Easier: Spin columns make DNA purification easier and more reliable - no more messy phenol-chloroform extractions.
      Greater Reproducibility: Positive and negative control antibodies and PCR primers are included to help validate your results and to troubleshoot your experiments.
      Alternate Names
      • Magnetic ChIP Kit
      Materials Required but Not DeliveredMagna Grip™ Rack 8 well (Cat.# 20-400) (Now Available!) or similar magnetic rack.
      Background InformationChromatin Immunoprecipitation (ChIP) is a powerful technique for mapping the in vivo distribution of proteins associated with chromosomal DNA. These proteins can be histone subunits and post-translational modifications or other chromatin associated proteins such as transcription factors, chromatin regulators, etc. Additionally, ChIP can be used to identify regions of the genome associated with these proteins, or conversely, to identify proteins associated with a particular region of the genome. ChIP methodology often involves protein-DNA and protein-protein cross-linking, fragmentation of the cross-linked chromatin, and subsequent immunoprecipitation of chromatin with an antibody specific to a target protein. The DNA fragments isolated in complex with the target protein can be identified by a variety of methods including PCR, DNA microarray and DNA sequencing. Standard or quantitative PCR can be performed to verify whether a particular DNA sequence (the gene or region of the genome) is associated with the protein of interest. The combination of ChIP and promoter or genomic tiling microarrays (ChIP-chip) allows genome-wide identification of DNA-binding sites for chromatin-associated proteins with precise resolution. Alternatively, high-throughput sequencing of libraries constructed from immunoprecipitated chromosomal DNA (ChIP-Seq) is a powerful alternative to ChIP-chip in mapping the protein-DNA interactions across mammalian genomes.
      References
      Product Information
      Components
      • Magnetic Protein A beads
      • ChIP Dilution Buffer
      • Low Salt Wash Buffer
      • High Salt Wash Buffer
      • LiCl Wash Buffer
      • TE Buffer
      • Cell Lysis Buffer
      • Nuclear Lysis Buffer
      • ChIP Elution Buffer (w/o Proteinase K)
      • 10X Glycine
      • 10X PBS
      • Protease Inhibitor Cocktail II
      • Proteinase K
      • Control Primers
      • Anti-Acetyl Histone H3
      • Normal Rabbit IgG
      • Spin Filters
      • Collection Tubes
      • Bind Reagent A
      • Wash Reagent B
      • Elution Reagent C
      PresentationTwo boxes containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions. Supplied buffers are sufficient to generate chromatin from up to five 15 cm plates of cultured cells, each plate providing up to 10 chromatin preparations (varies with cell and assay type).
      Applications
      ApplicationSingle day chromatin immunoprecipitation (ChIP) kit containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using magnetic A beads. Control primers included.
      Biological Information
      Analytes Available
      • Protein A
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsUpon receipt, store components at the temperatures indicated on the labels. Kit components are stable for 1 year from date of shipment when stored as directed.
      Packaging Information
      Material Size22 assays
      Material PackageKit capacity: 22 chromatin immunoprecipitation assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      EZ-Magna ChIP™ A - Chromatin Immunoprecipitation Kit MSDS

      타이틀

      물질안전보건자료(MSDS) 

      EZ-Magna ChIP™ A - Chromatin Immunoprecipitation Kit Certificates of Analysis

      TitleLot Number
      EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - DAM1421807DAM1421807
      EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - DAM1432915DAM1432915
      EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - DAM1439047DAM1439047
      EZ-Magna ChIPtrade; A Chromatin Immunoprecipitation Kit - R0708G0031R0708G0031
      Magna ChIP A or EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - 19826421982642
      Magna ChIP A or EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - 20145332014533
      Magna ChIP A or EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - 20323162032316
      Magna ChIP A or EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - 20456922045692
      Magna ChIP A or EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - 20890082089008
      Magna ChIP A or EZ-Magna ChIP A Chromatin Immunoprecipitation Kits - 21370042137004

      References

      Reference overviewPub Med ID
      Therapeutic potential of GSK-J4, a histone demethylase KDM6B/JMJD3 inhibitor, for acute myeloid leukemia.
      Li, Y; Zhang, M; Sheng, M; Zhang, P; Chen, Z; Xing, W; Bai, J; Cheng, T; Yang, FC; Zhou, Y
      J Cancer Res Clin Oncol  144  1065-1077  2018

      요약 표시
      29594337 29594337
      The Human Cytomegalovirus Strain DB Activates Oncogenic Pathways in Mammary Epithelial Cells.
      Kumar, A; Tripathy, MK; Pasquereau, S; Al Moussawi, F; Abbas, W; Coquard, L; Khan, KA; Russo, L; Algros, MP; Valmary-Degano, S; Adotevi, O; Morot-Bizot, S; Herbein, G
      EBioMedicine  30  167-183  2018

      요약 표시
      29628341 29628341
      Shifts in podocyte histone H3K27me3 regulate mouse and human glomerular disease.
      Majumder, S; Thieme, K; Batchu, SN; Alghamdi, TA; Bowskill, BB; Kabir, MG; Liu, Y; Advani, SL; White, KE; Geldenhuys, L; Tennankore, KK; Poyah, P; Siddiqi, FS; Advani, A
      J Clin Invest  128  483-499  2018

      요약 표시
      29227285 29227285
      Restoring Tip60 HAT/HDAC2 Balance in the Neurodegenerative Brain Relieves Epigenetic Transcriptional Repression and Reinstates Cognition.
      Panikker, P; Xu, SJ; Zhang, H; Sarthi, J; Beaver, M; Sheth, A; Akhter, S; Elefant, F
      J Neurosci  38  4569-4583  2018

      요약 표시
      29654189 29654189
      Myeloid-Specific Gene Deletion of Protein Phosphatase 2A Magnifies MyD88- and TRIF-Dependent Inflammation following Endotoxin Challenge.
      Sun, L; Pham, TT; Cornell, TT; McDonough, KL; McHugh, WM; Blatt, NB; Dahmer, MK; Shanley, TP
      J Immunol  198  404-416  2017

      요약 표시
      27872207 27872207
      Early growth response-1-mediated down-regulation of drebrin correlates with loss of dendritic spines.
      Cho, C; MacDonald, R; Shang, J; Cho, MJ; Chalifour, LE; Paudel, HK
      J Neurochem  142  56-73  2017

      요약 표시
      28369888 28369888
      Epigenetic mechanisms underlying NMDA receptor hypofunction in the prefrontal cortex of juvenile animals in the MAM model for schizophrenia.
      Gulchina, Y; Xu, SJ; Snyder, MA; Elefant, F; Gao, WJ
      J Neurochem  143  320-333  2017

      요약 표시
      28628228 28628228
      miR-130b directly targets ARHGAP1 to drive activation of a metastatic CDC42-PAK1-AP1 positive feedback loop in Ewing sarcoma.
      Satterfield, L; Shuck, R; Kurenbekova, L; Allen-Rhoades, W; Edwards, D; Huang, S; Rajapakshe, K; Coarfa, C; Donehower, LA; Yustein, JT
      Int J Cancer  141  2062-2075  2017

      요약 표시
      28748534 28748534
      Epigenetic Regulation of Cytosolic Phospholipase A2 in SH-SY5Y Human Neuroblastoma Cells.
      Tan, CS; Ng, YK; Ong, WY
      Mol Neurobiol  53  3854-3872  2016

      요약 표시
      26162318 26162318
      Tip60 HAT Action Mediates Environmental Enrichment Induced Cognitive Restoration.
      Xu, S; Panikker, P; Iqbal, S; Elefant, F
      PLoS One  11  e0159623  2016

      요약 표시
      27454757 27454757

      Brochure

      Title
      Shaping Epigenetics Discovery - Epigenetics Product Selection Brochure

      Technical Info

      Title
      White Paper - The Message in the Marks: Deciphering Cancer Epigenetics

      Data Sheet

      Title
      Reprogramming Cell Fate and Function Novel Strategies for iPSC Generation, Characterization, and Differentiation

      FAQ

      QuestionAnswer
      How should I resuspend my pellet prior to PCR?You should resuspend your pellet in water and not TE as the EDTA found in the TE may interfere with PCR.
      How many PCR reactions can be done with this kit?There are enough primers and PCR buffer for 4 reactions per IP assuming a 20 microliter volume and assuming the primers are at the recommended concentration as stated in the manual.
      Is there ever a time when I do not need to cross-link Histones?In native ChIP, Histone H3 and Histone H4 do not need to be crosslinked as they are very tightly associated. Histone H2A and Histone H2B are not as tightly associated, but will still work in native ChIP.
      From where are the primer sequences derived for the kit?The primer sequences are based on the Human GAPDH promoter. The GenBank number is NT_009759.15, using nts:6497145-6498136.
      What were your conditions for PCR?Please see the manual for The EZ ChIP Kit (Catalog #17-371) for more information.
      If I wanted to quantitate my immunoprecipitated DNA, how would I do so?DNA purified from ChIP experiments can be quantitated by PCR, providing the amplifying oligos meet specific criteria. Oligos should be 24 mers, with a GC content of 50% (+/- 4) and a Tm of 60.0C (+/- 2.0). You must be certain that the PCR reactions are within the linear range of amplification. Generally it takes time to achieve this. Too much input DNA will affect your results, so set up several tubes for each experiment to optimize the input DNA. Generally, this is about 1/25th to 1/100th for yeast, approximately 1/10 for mammalian cells, but depends on the amount of antibody and input chromatin.

      Also, do not use more than 20 cycles, making sure that dNTP's always remain in excess. Also, include each reaction a control primer (to compare your experimental band against-make sure the sizes are sufficiently different to allow proper separation-75 base pairs is usually OK) set to a region of the genome that should not change throughout your experimental conditions. Also PCR from purified input DNA (no ChIP) and include no antibody control PCR's as well. PCR products should be no more than 500 base pairs and should span the area of interest (where you think you will see changes in acetylation or methylation of histones). All PCR products should be run on 7-8% acrylamide gels and stained with SYBR Green 1 (Molecular Probes) at a dilution of 1:10,000 (in 1X Tris-borate-EDTA buffer, pH 7.5) for 30 minutes-no destaining is required.

      Quantitation is carried out subsequent to scanning of the gel on a Molecular Dynamics Storm 840 or 860 in Blue fluorescence mode with PMT voltage at 900 with ImageQuant software. This has distinct advantages over ethidium bromide staining. SYBR Green is much more sensitive, and illumination of ethidium stained gels can vary across the gel based on the quality of UV bulbs in your in your light box. For further info, see Strahl-Bolsinger et al. (1997) Genes Dev. 11: 83-93. A radioactive quantitation m
      I am not getting amplification with input DNA. What did I do wrong?Your input DNA sample should be taken just prior to adding the antibody. It is considered the starting material. If you are not seeing amplification with your input DNA, either you have not successfully reversed the cross links or the PCR is not working for reasons other than the kit.
      Do you have any tips for sonication?Keep cells on ice throughout the procedure - even during sonication. Be sure that you don't sonicate for to long (more than 30 seconds could cause sample overheating and denaturation).
      Why is more DNA is precipitated in my no-antibody control than for my test sample?To eliminate banding in your negative controls you can do several things:

      A) Pre-clear the 2ml diluted cell pellet suspension with 80 microliters of Salmon Sperm DNA/Protein A Agarose-50% Slurry for 30 minutes at 4ºC with agitation. You could try to preclear the lysate longer or with more clearings.

      B) Titrate your input DNA, to see when the bands in the NFA disappear.

      C) Use an alternative lysis procedure: Resuspend cell pellet in 200 microliters of 5mM Pipes pH 8.0, 85mM KCl, 0.5% NP40 containing protease inhibitors. Place on ice for 10 minutes. Pellet by centrifugation (5 minutes at 5000 rpm). Resuspend pellet in 200 microliters of 1% SDS, 10mM EDTA, 50mM Tris-HCl, pH 8.1 containing protease inhibitors. Incubate on ice for 10 minutes.

      D) Block the Salmon Sperm DNA Agarose prior to use in 1-5% BSA and Chip dilution buffer (mix at room temperature for 30 minutes). After incubation, spin the agarose and remove the 1% BSA/ChIP assay buffer supernatant. Wash once in ChIP assay buffer and continue.
      What is 'Input DNA', and why no 'Output DNA'?Input DNA is DNA obtained from chromatin that has been cross-link reversed similar to your samples. It is a control for PCR effectiveness. Output DNA is the DNA from each of your ChIP experiments.

      User Guides

      Title
      EZ-Magna ChIP™ A

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      카테고리

      Life Science Research > Kits & Assays > ChIP Kits > Kits and Assays
      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Kits & Beads