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  1. Link

    rad21-antibody-chip-grade-ab992.pdf

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Cell Biology Cell Cycle Cell Division Chromatid Cohesion
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Anti-Rad21 抗体 - ChIP Grade (ab992)

  • Datasheet
  • SDS
Reviews (19)Q&A (9)References (109)

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  • Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody - ChIP Grade (ab992)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody - ChIP Grade (ab992)
  • Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
  • Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)

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関連製品

  • データシート
  • 文献 (109)
  • プロトコール

製品の概要

  • 製品名

    Anti-Rad21 antibody - ChIP Grade
    Rad21 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Rad21 - ChIP Grade
  • 由来種

    Rabbit
  • 特異性

    The epitope recognized by ab992 maps to a region between residue 575 and the C-terminus (residue 631) human Rad21 homolog using the numbering given in entry NP_006256.1 (GeneID 5885).
  • アプリケーション

    適用あり: IHC-P, ChIP, ICC/IF, WB, IPmore details
  • 種交差性

    交差種: Mouse, Human, Xenopus laevis, Indian muntjac
    交差が予測される動物種: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Turkey, Chimpanzee, Gorilla, Chinese hamster, Orangutan, Elephant
  • 免疫原

    Synthetic peptide (Human) conjugated to KLH - which represented a portion of human Rad21 encoded within exon 14 (LocusLink ID 5885).

  • ポジティブ・コントロール

    • HeLa cell lysate. FFPE human breast fibroadenoma tissue sections.
  • 特記事項

      

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファー

    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Chromatid Cohesion
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • Other
    • Epigenetics and Nuclear Signaling
    • Chromosome Structure
    • Cohesins
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Chromosome Structure
    • Chromatid Cohesion
    • Cancer
    • Cell cycle
    • Cell division
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

関連製品

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Rad21 protein (ab152647)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

アプリケーション

Our Abpromise guarantee covers the use of ab992 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Used at a dilution of 1/200 for 30 min incubation (see Abreview for further information).

WB 1/500 - 1/5000. Predicted molecular weight: 72 kDa.

Band observed at ~130 kDa.

IP Use a concentration of 1 - 4 µg/ml.

ターゲット情報

  • 機能

    Cleavable component of the cohesin complex, involved in chromosome cohesion during cell cycle, in DNA repair, and in apoptosis. The cohesin complex is required for the cohesion of sister chromatids after DNA replication. The cohesin complex apparently forms a large proteinaceous ring within which sister chromatids can be trapped. At metaphase-anaphase transition, this protein is cleaved by separase/ESPL1 and dissociates from chromatin, allowing sister chromatids to segregate. The cohesin complex may also play a role in spindle pole assembly during mitosis. Also plays a role in apoptosis, via its cleavage by caspase-3/CASP3 or caspase-7/CASP7 during early steps of apoptosis: the C-terminal 64 kDa cleavage product may act as a nuclear signal to initiate cytoplasmic events involved in the apoptotic pathway.
  • 配列類似性

    Belongs to the rad21 family.
  • ドメイン

    The C-terminal part associates with the head of SMC1A, while the N-terminal part binds to the head of SMC3.
  • 翻訳後修飾

    Cleaved by separase/ESPL1 at the onset of anaphase. Cleaved by caspase-3 and caspase-7 at the beginning of apoptosis. The cleavage by ESPL1 and caspase-3 take place at different sites.
    Phosphorylated; becomes hyperphosphorylated in M phase of cell cycle. The large dissociation of cohesin from chromosome arms during prophase may be partly due to its phosphorylation by PLK.
  • 細胞内局在

    Nucleus. Chromosome. Chromosome > centromere. Associates with chromatin. Before prophase it is scattered along chromosome arms. During prophase, most of cohesin complexes dissociate from chromatin probably because of phosphorylation by PLK, except at centromeres, where cohesin complexes remain. At anaphase, it is cleaved by separase/ESPL1, leading to the dissociation of the complex from chromosomes, allowing chromosome separation. Once cleaved by caspase-3, the C-terminal 64 kDa cleavage product translocates to the cytoplasm, where it may trigger apoptosis.
  • Target information above from: UniProt accession O60216 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 540966 Cow
    • Entrez Gene: 5885 Human
    • Entrez Gene: 19357 Mouse
    • Entrez Gene: 314949 Rat
    • Entrez Gene: 399129 Xenopus laevis
    • Omim: 606462 Human
    • SwissProt: Q3SWX9 Cow
    • SwissProt: O60216 Human
    • SwissProt: Q61550 Mouse
    • SwissProt: O93310 Xenopus laevis
    • Unigene: 81848 Human
    • Unigene: 182628 Mouse
    • Unigene: 3991 Rat
    • Unigene: 53864 Xenopus laevis
    see all
  • 別名

    • CDLS4 antibody
    • Double-strand-break repair protein rad21 homolog antibody
    • hHR21 antibody
    • HR21 antibody
    • HRAD21 antibody
    • KIAA0078 antibody
    • MCD1 antibody
    • Nuclear matrix protein 1 antibody
    • NXP-1 antibody
    • NXP1 antibody
    • Protein involved in DNA double-strand break repair antibody
    • RAD21 antibody
    • RAD21 homolog (S. pombe) antibody
    • RAD21 homolog antibody
    • RAD21_HUMAN antibody
    • Scc1 antibody
    • SCC1 homolog antibody
    see all

画像

  • Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
    Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
    Anti-Rad21 antibody - ChIP Grade (ab992) at 1/1000 dilution + HeLa whole cell lysate

    Developed using the ECL technique.

    Predicted band size: 72 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 130 kDa (possible cross reactivity)

  • Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody - ChIP Grade (ab992)
    Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody - ChIP Grade (ab992)

    ab992 at a 1/200 dilution staining Asynchronous and paraformaldehyde-fixed (4%) HeLa cells by immunocytochemistry. The antibody was incubated with the cells 30 minutes and then detected using a Cy3 conjugated Goat Anti-Mouse IgG (H+L) antibody.

    This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody - ChIP Grade (ab992)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody - ChIP Grade (ab992)
    IHC image of Rad21 staining in human breast fibroadenoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab992, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
    Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
    Anti-Rad21 antibody - ChIP Grade (ab992) at 1/2000 dilution + HeLa whole cell extract (extraction was achieved using RIPA buffer and 1% SDS) at 25 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/1500 dilution

    Predicted band size: 72 kDa



    A rad21 knock down negative control was employed.

    This image is courtesy of an Abreview submitted on 9 September 2005. We do not have any further information relating to this image.

    See Abreview

  • Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
    Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)
    All lanes : Anti-Rad21 antibody - ChIP Grade (ab992) at 2 µg/ml

    Lane 1 : HeLa at 50 µg
    Lane 2 : HeLa at 15 µg
    Lane 3 : HeLa at 5 µg
    Lane 4 : 293T at 50 µg
    Lane 5 : NIH3T3 at 50 µg

    Predicted band size: 72 kDa


    Exposure time: 30 seconds

プロトコール

  • ChIP protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

データシートおよび資料

    • Datasheet
    • SDS
  • 参考文献

    This product has been referenced in:

    • Amat R  et al. Rapid reversible changes in compartments and local chromatin organization revealed by hyperosmotic shock. Genome Res 29:18-28 (2019). Read more (PubMed: 30523037) »
    • Meisenberg C  et al. Repression of Transcription at DNA Breaks Requires Cohesin throughout Interphase and Prevents Genome Instability. Mol Cell 73:212-223.e7 (2019). Read more (PubMed: 30554942) »
    See all 109 Publications for this product

    ab992 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 28 Abreviews or Q&A

    Question

    Customer kindly contacted us regarding positve and negative primers for our ChIP grade antibodies.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 06 2013

    Answer

    I've compiled some information below regarding potential primer sets for the ChIP antibodies we talked about, where information was lacking (verified in ChIP by other researches), I included links to abReviews and references that used the antibody in ChIP.

    1. ab817: GAPDH (+), GAD1 (-), AFM (-), Factor VIII (-)

    2. ab1791: GAPDH, RPL30, ALDOA, MYO-D, SERPINA, AFM, SAT2, SATa (each batch is verified against this primer set, an example image of one batch test is given on the datasheet)

    3. ab7030: was verified in ChIP by other researchers submitting data through our abReview system and specific references. Their positive and negative controls used are given through the link below, as well as a "click to contact this user" link that allows you to contact the researchers if you would like more specific primer information. The specific references with ChIP listed in bold are good sources for positive and negative primers as well.
    a) Abreview: (NFKBIA promoter (+), GAPDH (-)) https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=25057
    b) Abreview: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=18386
    c) Specific references: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-references.html

    4. ab111069 Untested in ChIP.

    5. ab992: was verified in ChIP by other researchers (abReviews and specific references)
    a) Specific references: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-references.html
    b) Abreview: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=14542
    c) Abreview: (EBV 3+ (+), EBV1,2,4,5,6 (-), GAPDH (-), Cen (-)) https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=33942

    Our ChIP primer sets:
    https://www.abcam.com/index.html?pageconfig=resource&rid=10600

    Read More

    Abcam Scientific Support

    Answered on Feb 06 2013

    ChIP abreview for Anti-Rad21 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (MCF10a)
    Negative control
    Random regions without ATAC-seq signal or any histone marks.
    Specification
    MCF10a
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde
    Positive control
    Strong CTCF binding sites
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 10 2019

    ChIP abreview for Anti-Rad21 antibody - ChIP Grade

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Mouse Tissue lysate - nuclear (Limb buds, forebrain, ESC)
    Negative control
    We perform ChIP-seq
    Specification
    Limb buds, forebrain, ESC
    Detection step
    Other
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 37% Formaldehyde (Sigma), dilu
    Positive control
    We perform ChIP-seq
    Read More

    Daniel Ibrahim

    Verified customer

    投稿 Nov 12 2018

    Western blot abreview for Anti-Rad21 antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Whole cell lysate resolved on a 12% SDS PAGE gel)
    Gel Running Conditions
    Reduced Denaturing (12% SDS gel)
    Loading amount
    25 µg
    Treatment
    HEK293T cells transfected with HA-Rad21 plasmid for overexpression
    Specification
    Whole cell lysate resolved on a 12% SDS PAGE gel
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 19 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad21 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Cells are incubated with primary antibody + BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
    Sample
    Donkey Cell (Skin fibroblasts)
    Specification
    Skin fibroblasts
    Permeabilization
    Yes - 1% PFA + 0.2% Triton
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 12 2014

    Question

    What does this antibody recognize on human Rad21?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 24 2013

    Answer

    The epitope recognized by ab992 maps to a region between residue 575 and the C-terminus (residue 631) human Rad21 homolog using the numbering given in entry NP_006256.1 (GeneID 5885).

    Read More

    Tom Ruyle

    Abcam Scientific Support

    Answered on Oct 24 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad21 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Sample
    Mouse Cell (primary hepatoctyes)
    Specification
    primary hepatoctyes
    Permeabilization
    Yes - Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
    Fixative
    Paraformaldehyde
    Read More

    Mr. Shengyun Ma

    Verified customer

    投稿 Oct 07 2013

    Immunoprecipitation abreview for Anti-Rad21 antibody - ChIP Grade

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein A
    Sample
    Human Cell lysate - whole cell (Hep3B)
    Specification
    Hep3B
    Total protein in input
    30 µg
    Read More

    Abcam user community

    Verified customer

    投稿 Sep 16 2013

    ChIP abreview for Anti-Rad21 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Detection step
    Real-time PCR
    Sample
    Mouse Cell lysate - whole cell (primary hepatoctyes)
    Specification
    primary hepatoctyes
    Negative control
    IgG
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% formaldehyde
    Positive control
    Suz12 on h19 ICR
    Read More

    Abcam user community

    Verified customer

    投稿 Sep 16 2013

    Western blot abreview for Anti-Rad21 antibody - ChIP Grade

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Non-reduced Denaturing (8%)
    Sample
    Mouse Cell lysate - whole cell (Primary hepatoctyes)
    Specification
    Primary hepatoctyes
    Blocking step
    I-Block(Applied biosystems) as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 4°C
    Read More

    Mr. Shengyun Ma

    Verified customer

    投稿 Aug 09 2013

    1-10 of 28 Abreviews or Q&A

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