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 제품안내  항체,항원,킷트   Cancer Research  Epigenetics

CUTANA® CUT&RUN Assays & Antibodies

ChIP-Seq은 타겟단백질이 결합된 크로마틴 부위를 침전시켜 농축하고 그 시퀀스를 NGS로 분석하는 기술입니다. 그러나 ChIP-Seq은 세포의 양이 적은 샘플의 경우 DNA를 얻기 힘들고, 노이즈 백그라운드가 높다는 한계를 갖고 있습니다.

CUT&RUN 시퀀싱은 온전한 세포로부터 타겟 단백질만을 잘라내는 방식으로, ChIP-Seq에 비해 적은 수의 cell 로도 분석이 가능하고, 시간이 단축되며, signal to noise ratio가 월등히 향상되어, Chromatin NGS에 효과적으로 이용될 수 있는 툴 입니다.

EpiCypher
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  • Histone 변형 및 Protein-DNA 상호 작용을 분석하고, 결합부위의 유전자를 규명하기 위해 이용됩니다.
  • 기존 ChIP, ChIP-seq의 민감도와 노이즈 문제를 보완하였습니다.
  • Intact cell이나 핵내에 타겟에 대한 항체를 반응시킨 후, Protein A-MNase fusion protein을 이용하여 타겟만을 특이적으로 절단하므로 백그라운드가 감소됩니다.
  • 적은양의 샘플(5,000개-500,000개 세포), 적은양의 sequencing reads로도 고품질의 시퀀싱 데이터를 얻을 수 있습니다.
  • Kit의 구성 : buffers, enzymes, magnetic beads, control antibodies, spike-in controls, 8-strip tubes, spin columns  
  • CUT&RUN assay에 이용할 수 있는 PTM 항체 및 크로마틴 리모델링 항체 (CHD1, SNF2L, SNF2H, BRM)가 출시되었습니다. (아래 항체 부분에서 자세한 사항을 확인하세요.)

 


     1) lectin-coated 된 magnetic bead에 cell을 고정하고 permeabilization화 합니다.
     2) 크로마틴 타겟(ex: 히스톤 PTM 또는 크로마틴/DNA 결합 단백질)에 대한 항체를 반응시킵니다.
     3) CUTANA pAG-MNase를 처리하여 타겟 nucleosome을 잘라냅니다.
     4) CaCl2+를 넣으면 nucleosome complex가 유리됩니다.
     5) nucleosome complex로부터 DNA를 extraction하여 sequencing library를 준비합니다.
     6) NGS를 진행합니다.







              * Assay 방법별 차이점 보기










      Comparing data generated from standard ChIP-seq methods and sequencing depth (black) to CUTANA (blue).
      H3K27me3 and H3K4me3 ChIP-Seq data was sourced from ENCODE. Negative control data (yellow) was 
      generated using a Rabbit IgG antibody in CUTANA. Each dataset of CUTANA was acquired using 
      0.5 million cells.





  • Skene and Henikoff. 2017. eLife.
  • Skene et al. 2018. Nat. Protocols.
  • Janssens et al. 2018. Epigenetics and Chromatin.
  • Thakur and Henikoff. 2018. Genes & Dev.
  • Brahma and Henikoff. 2019. Mol. Cell.
  • Meers et al. 2019. eLife.
  • Akhtar et al. 2019. Life Sci Alliance.
  • de Bock et al. 2018. Cancer Discovery.
  • Ernst et al. 2019. Nature Comm.
  • Hainer and Fazzio. 2019. Current Prot.in Mol. Bio.
  • Hainer et al. 2019. Cell.
  • Hyle et al. 2019. Nucleic Acids Res.
  • Inoue et al. 2018. Genes & Dev.
  • Liu et al. 2018. Cell.
  • Menon et al. 2019. Development.
  • Oomen et al. 2019. Genome Res.
  • Park et al. 2019. Cell Stem Cell.
  • Roth et al. 2018. Nature.
  • Uyehara and McKay. 2019. PNAS.
  • Zheng and Gehring. 2019. Plant Reprod.
  • Mathsyaraja H et al. 2019. G&D

 







  CUTANA™ ChIC/CUT&RUN Kit - 48 rxns (cat# 14-1048)
  • CUT&RUN Assay에 필요한 모든 시약 제공
  • multi-channel pipette으로 사용하기 편리하도록 48 샘플 테스트용으로 제공
  • H3K4me3 (positive control)과 IgG (negative control)을 함께 제공하여 실험결과의 정확성을 입증
  • cell, nuclei sample에 적용가능
  • 5000개 정도의 적은양의 cell로도 실험 가능
  • Compatible with a variety of targets: histone PTMs, transcription factors, epigenetic enzymes and reader proteins, chromatin remodeling proteins
  • Validated protocols for diverse sample preparations: cells, nuclei, cryopreserved and crosslinked
  • 제품의 구성 : buffers, enzymes, 8-strip tubes, magnetic beads, positive/negative control antibodies, CUTANA™ dNuc spike-in controls (for experimental troubleshooting), E. coli Spike-in DNA (for data normalization), and DNA purification spin columns

  • CUT&RUN DNA Fragment Size Distribution Analysis. CUT&RUN was performed using the CUTANA ChIC/CUT&RUN Kit starting with 500,000 K562 cells. CUT&RUN DNA isolated from IgG Negative Control (13-0042k) and H3K4me3 Positive Control (13-0041k) antibodies was used to prepare paired-end Illumina sequencing libraries. Library DNA was analyzed by Agilent Tapestation®. This analysis confirmed that mononucleosomes  were predominantly enriched in CUT&RUN (~300 bp peak represents 150 bp nucleosomes +  sequencing adapters).


  • CUTANA H3K4 MetStat Spike-in Controls. DNA-barcoded unmodified and H3K4-methylated dNucs were immobilized to Streptavidin Beads and spiked-in to CUT&RUN samples prior to the addition of either IgG (top) or H3K4me3 (bottom) control antibodies. The shell script available on the product page was used to count instances of each barcoded dNuc in the CUT&RUN sequencing data. The proportion of read counts normalized to on-target (H3K4me3) are shown. The spike-ins confirmed that the control antibody specifically recovered the target dNuc.
  • View technical datasheet for this product. 14-1048 Datasheet
  • View Kit Manual. CUTANA ChIC/CUT&RUN Kit Manual
  • View Kit Quick Card. CUTANA ChIC/CUT&RUN Quick Card
  • H3K4 MetStat Spike-in Controls Analysis H3K4 MetStat Spike-in Controls Analysis_v1
  • View H3K4 MetStat Spike-in Controls Shell Script CUTANA H3K4 MetStat Spike-in Controls Shell Script

 

  CUTANA® pAG-MNase for ChIC/CUT&RUN Workflows - 50 rxns (cat# 15-1016)
  • Type:  Nuclease
  • Host: E.coli
  • Mol Wgt: 44 kDa
  • Epitope Tag: 6His
  • Description : Recombinantly produced in E. coli, CUTANA™ pAG-MNase for ChIC/CUT&RUN Workflows is a fusion of Proteins A and G to Micrococcal Nuclease. This construct is useful in performing Chromatin Immunocleavage (ChIC) and Cleavage Under Targets and Release Using Nuclease (CUT&RUN). CUTANA™ pAG-MNase contains a C-terminal 6xHis epitope tag.
  • Formulation : provided as a 20X stock in 10 mM Tris HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, and 50% glycerol.
  • Application : This product is sufficient to perform 50 CUT&RUN reactions. Recommended use: 2.5 μL of the supplied enzyme into a 50 μL CUT&RUN reaction (20X dilution). 
  CUTANA™ DNA Purification Kit - 50 samples (cat# 14-0050)
  • CUT&RUN을 통해 얻은 DNA fragment를 정제하기 위한 kit
  • 30분만에 정제된 농축 DNA 준비 완료
  • 컬럼을 이용하여 capture & recovery가 높음

  • CUT&RUN DNA Fragment Size Distribution Analysis. CUT&RUN was performed using the CUTANA ChIC/CUT&RUN Kit starting with 500,000 K562 cells. CUT&RUN DNA isolated from IgG negative control (EpiCypher Catalog No. 13-0042), H3K4me3 (EpiCypher Catalog No. 13-0041), H3K27me3 (EpiCypher Catalog No. 13-0030) and CTCF (EMD Millipore Catalog No. 07-729) antibodies were used to prepare paired-end Illumina sequencing libraries. Library DNA was analyzed by Agilent Tapestation®. This analysis confirmed that mononucleosomes  were predominantly enriched in CUT&RUN (~300 bp peak represents 150 bp nucleosomes +  sequencing adapters). 
  • View technical datasheet for this product. 14-0050 Datasheet
  • View Kit Manual. CUTANA ChIC/CUT&RUN Kit Manual

 

  CUTANA® E. coli Spike-in DNA - 50 rxns (cat# 18-1401)
  • Data normalization을 위한 E. coli DNA 제공
  • CUT&RUN 실험 시 Exogenous E. coli DNA를 주입해 DNA Purification 및 library sequencing 준비를 control 
  CUTANA® CUT&RUN Compatible Antibodies
  • 13-0042 Rabbit IgG Antibody, CUTANA®  CUT&RUN, Negative Control
  • 13-0041 H3K4me3 Antibody: SNAP-ChIP® Certified, CUTANA® CUT&RUN Compatible
  • 13-0030 H3K27me3 Antibody: SNAP-ChIP® Certified, CUTANA® CUT&RUN Compatible
  • 13-0031 H3K36me3 Antibody: SNAP-ChIP® Certified, CUTANA® CUT&RUN Compatible 
  • 13-2004 MLL1/KMT2A CUTANA CUT&RUN Antibody (NEW / 2021 Feb)
  • 13-2005 SNF2L/SMARCA1 CUTANA CUT&RUN Antibody (NEW / 2021 Feb)
  • 13-2006 BRM/SMARCA2 CUTANA CUT&RUN Antibody (NEW / 2021 Feb)
  • 13-2007 SNF2H/SMARCA5 CUTANA CUT&RUN Antibody (NEW / 2021 Feb)
  • 13-2008 CHD1 CUTANA CUT&RUN Antibody (NEW / 2021 Feb)

 







  CUTANA® Tools
  • 10-0008 Magnetic Separation Rack, 16 x 0.2ml tube
  • 10-0012 Magnetic Separation Rack, 6 x 1.5ml tube
  • 21-1401 CUTANA®  Concanavalin A-Conjugated Paramagnetic Beads, 550ul
  • 21-1411 CUTANA®  Concanavalin A-Conjugated Paramagnetic Beads, 2.75ml
  • 10-0009 8-strip 0.2mL PCR strip tubes, 120 strips