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  1. Link

    rest--nrsf-antibody-ab21635.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Zinc Finger
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Anti-REST / NRSF 抗体 (ab21635)

  • Datasheet
  • SDS
Reviews (4)Q&A (2)References (29)

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Western blot - Anti-REST / NRSF antibody (ab21635)

    Key features and details

    • Rabbit polyclonal to REST / NRSF
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

    こちらの製品もご検討ください

    一次抗体
    Product image
    Anti-5 Lipoxygenase/5-LO antibody [EP6072(2)] (ab169755)
    二次抗体
    Product image
    Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    ノックアウト
    Product image
    Human REST (NRSF) knockout HeLa cell line (ab265017)

    関連製品

    製品の概要

    • 製品名

      Anti-REST / NRSF antibody
      REST / NRSF 一次抗体 製品一覧
    • 製品の詳細

      Rabbit polyclonal to REST / NRSF
    • 由来種

      Rabbit
    • アプリケーション

      適用あり: WBmore details
      適用なし: IHC-P
    • 種交差性

      交差種: Human
      交差が予測される動物種: Mouse, Rat
    • 免疫原

      Synthetic peptide corresponding to Mouse REST/ NRSF aa 1050 to the C-terminus (internal sequence) conjugated to keyhole limpet haemocyanin.
      (Peptide available as ab17042)

    • 特記事項

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    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 or -80°C. Avoid freeze / thaw cycle.
    • バッファー

      pH: 7.40
      Preservative: 0.02% Sodium azide
      Constituent: PBS

      Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
    • 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

      • Epigenetics and Nuclear Signaling
      • Transcription
      • Domain Families
      • Zinc Finger
      • Neuroscience
      • Neurology process
      • Neurogenesis
      • Epigenetics and Nuclear Signaling
      • Transcription
      • Co-factors

    関連製品

    • Compatible Secondaries

      • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
      • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Immunizing Peptide (Blocking)

      • Mouse REST / NRSF peptide (ab17042)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
    • KO cell lines

      • Human REST (NRSF) knockout HeLa cell line (ab265017)
    • KO cell lysates

      • Human REST (NRSF) knockout HeLa cell lysate (ab257636)

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab21635の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    WB (2)
    Use a concentration of 1 µg/ml. Detects a band of approximately 121 kDa (predicted molecular weight: 121 kDa).
    特記事項
    WB
    Use a concentration of 1 µg/ml. Detects a band of approximately 121 kDa (predicted molecular weight: 121 kDa).
    追加情報
    Is unsuitable for IHC-P.

    ターゲット情報

    • 機能

      Transcriptional repressor which binds neuron-restrictive silencer element (NRSE) and represses neuronal gene transcription in non-neuronal cells. Restricts the expression of neuronal genes by associating with two distinct corepressors, mSin3 and CoREST, which in turn recruit histone deacetylase to the promoters of REST-regulated genes. Mediates repression by recruiting the BHC complex at RE1/NRSE sites which acts by deacetylating and demethylating specific sites on histones, thereby acting as a chromatin modifier.
    • 組織特異性

      Ubiquitous. Expressed at higher levels in the tissues of the lymphocytic compartment, including spleen, thymus, peripheral blood lymphocytes and ovary.
    • 配列類似性

      Contains 9 C2H2-type zinc fingers.
    • 細胞内局在

      Nucleus.
    • Target information above from: UniProt accession Q13127 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: 5978 Human
      • Entrez Gene: 19712 Mouse
      • Entrez Gene: 83618 Rat
      • Omim: 600571 Human
      • SwissProt: Q13127 Human
      • SwissProt: Q155C6 Mouse
      • SwissProt: Q8VIG1 Mouse
      • SwissProt: O54963 Rat
      • SwissProt: O54964 Rat
      • Unigene: 307836 Human
      • Unigene: 28840 Mouse
      • Unigene: 482359 Mouse
      • Unigene: 10879 Rat
      see all
    • 別名

      • Neural Restrictive Silencer Factor antibody
      • Neural-restrictive silencer factor antibody
      • Neuron Restrictive Silencer Factor antibody
      • NRSF antibody
      • RE1 silencing transcription factor antibody
      • RE1-silencing transcription factor antibody
      • Repressor binding to the X2 box antibody
      • rest antibody
      • REST protein antibody
      • REST_HUMAN antibody
      • X2 box repressor antibody
      • XBR antibody
      see all

    画像

    • Western blot - Anti-REST / NRSF antibody (ab21635)
      Western blot - Anti-REST / NRSF antibody (ab21635)
      All lanes : Anti-REST / NRSF antibody (ab21635) at 1 µg/ml

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : Wild-type HeLa Vehicle Control DFO (0mM, 16h) Nuclear cell lysate
      Lane 3 : Wild-type HeLa Treated DFO (0.5mM, 16h) Nuclear cell lysate
      Lane 4 : REST knockout HeLa Vehicle Control DFO (0mM, 16h) Nuclear cell lysate
      Lane 5 : REST knockout HeLa Treated DFO (0.5mM, 16h) Nuclear cell lysate
      Lane 6 : ES-D3 Nuclear Fraction cell lysate

      Lysates/proteins at 40 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 121 kDa
      Observed band size: 180 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-REST / NRSF antibody staining at 1 ug/ml, shown in green; Mouse anti SIRT1 nuclear loading control staining at 1/20000 dilution, shown in red. In Western blot, ab21635 was shown to bind specifically to REST / NRSF. A band was observed at 180 kDa in wild-type HeLa cell lysates with no signal observed at this size in resT CRISPR-Cas9 edited cell line ab265017 (CRISPR-Cas9 edited cell lysate ab257636). The band observed in the CRISPR-Cas9 edited lysate lane below 180 kDa is likely to represent a truncated form of REST / NRSF. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and resT CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    プロトコール

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    データシートおよび資料

    • SDS download

    • Datasheet download

      Download

    参考文献 (29)

    ab21635 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。

    ab21635 は 29 報の論文で使用されています。

    • Bebber CM  et al. Ferroptosis response segregates small cell lung cancer (SCLC) neuroendocrine subtypes. Nat Commun 12:2048 (2021). PubMed: 33824345
    • Singh NP  et al. Genome-Wide Binding Analyses of HOXB1 Revealed a Novel DNA Binding Motif Associated with Gene Repression. J Dev Biol 9:N/A (2021). PubMed: 33546292
    • Rossi C  et al. Cell Stress Induces Mislocalization of Transcription Factors with Mitochondrial Enrichment. Int J Mol Sci 22:N/A (2021). PubMed: 34445555
    • Bray JK  et al. Loss of RE-1 silencing transcription factor accelerates exocrine damage from pancreatic injury. Cell Death Dis 11:138 (2020). PubMed: 32080178
    • Xie Y & Cheng Y LINC01198 facilitates gliomagenesis through activating PI3K/AKT pathway. RNA Biol 17:1040-1052 (2020). PubMed: 32378450
    View all Publications for this product

    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

    1-6 of 6 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-REST / NRSF antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HAFs)
    Permeabilization
    Yes - 0.2% TritonX100/PBS
    Specification
    HAFs
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 18 2022

    Western blot abreview for Anti-REST / NRSF antibody

    Inconclusive
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Rat Tissue lysate - nuclear (Hippocampus, liver and MCF7 whole cell homogenates)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    80 µg
    Specification
    Hippocampus, liver and MCF7 whole cell homogenates
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Octavio Mercado-Gomez

    Verified customer

    投稿 May 12 2016

    Western blot abreview for Anti-REST / NRSF antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    90 µg
    Gel Running Conditions
    Reduced Denaturing (4-18% gel)
    Sample
    Mouse Cell lysate - whole cell (Mouse embryonic stem cells)
    Specification
    Mouse embryonic stem cells
    Blocking step
    BSA 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 Mar 17 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-REST / NRSF antibody

    Poor
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    Yes - 0.3% TRITON-X
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Jul 14 2014

    Question

    LOT NUMBER GR3030 ORDER NUMBER 974618 DESCRIPTION OF THE PROBLEM No staining SAMPLE I used pancreas as tissue of interest together with spleen as one of the positive controls recomended by abcam PRIMARY ANTIBODY ab21635 used 1:100 and 1:500 (O/N at 4C) DETECTION METHOD IF POSITIVE AND NEGATIVE CONTROLS USED No primary antibody. ANTIBODY STORAGE CONDITIONS Aliquoted after arrival and kept at -20C FIXATION OF SAMPLE Tried frozen sections prefixed with PFA4% and cryopreserved with sucrose 30% before OCT embedding and not fixed frozen sections that where fixed after sectioning with cold methanol for 10 min. ANTIGEN RETRIEVAL No PERMEABILIZATION STEP yes, 15 min PBS-Triton 0.5% BLOCKING CONDITIONS PBS-TRITON 0.5% with 10% normal donkey serum SECONDARY ANTIBODY anti rabbit Alexafluor488 (1-2h at RT) at 1:200 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Concentration of primary antibody Time incubation secondary antibody Fixation method

    Read More

    Abcam community

    Verified customer

    Asked on Nov 10 2011

    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality. Reviewing this case, I would like to offer some suggestions to help optimise the results from ab21635. I would also appreciate if you can confirm some further details: 1. I am sorry to confirm this antibody has not previously been tested in Oct embedded sections. Have you tried in fresh frozen sections, not embedded, which are tested and guaranteed? I can recommend to try this alternative if possible. 2. I can suggest there is no need to fix in methanol, as the tissue is already fixed n 4% PFA. If you want to be sure, I can recommend to fix in 4% PFA for a further 10 minutes before proceeding with staining. 2. 0.5% is quite a high concentration of Triton. This is a strong detergent which may disrupt proteins. Try 0.1% for 10 minutes only. This will help permeabilize the cells and nucleus. 3. Could you confirm if the current vial of secondary antibody is working well with other primary antibodies. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    Abcam Scientific Support

    Answered on Nov 10 2011

    Question

    we recently received the anti-Rest/NRSF antibody (Ab21635). I am attaching Immunhistochemistry results on male meiotic sections here (mouse tissue). What can be seen is clearly different from what we know about Rest expression. Rest is expressed in the spermatogonia type 2 and also in Pachytene spermatocytes and it shows nuclear localization (transcription factor).  The staining we get is tubulin-like - what we see is actually the spindle aparatus. Either we obtained the wrong antibody, or something is wrong with this antibody. Looking forward to your response!

    Read More

    Abcam community

    Verified customer

    Asked on Oct 05 2011

    Answer

    Thank you for contacting us. I have forwarded your data to our lab for analysis. It would be very helpful if you could please provide some additional details about your protocol: 1.) How were the tissues fixed and processed (paraffin or frozen, PFA fixation, etc)? We have only validated this antibody for use on PFA-fixed frozen sections. 2.) Did you perform any antigen retrieval, permeabilization, or other pretreatments? 3.) How were the sections blocked? 4.) What were the primary and secondary antibody dilutions and incubation times? 5.) What secondary antibody was used? Did the no primary antibody control come out clean? 6.) Could you please provide your original order or lot number? I will be happy to assist you further if you could please provide this additional information.

    Read More

    Abcam Scientific Support

    Answered on Oct 05 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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