Anti-SMARCA6 抗体 (ab3851)

Reviews (3)Q&A (2)References (1)

Key features and details

  • Rabbit polyclonal to SMARCA6
  • Suitable for: WB, IP
  • Reacts with: Mouse, Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-SMARCA6 antibody

  • 製品の詳細

    Rabbit polyclonal to SMARCA6

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IPmore details

  • 種交差性

    交差種: Mouse, Human

  • 免疫原

    Synthetic peptide (Human) - which represents a portion of human Helicase Lymphoid Specific encoded in part by exons 15 and 16.

  • 特記事項


    DNA helicases (Hel) play a role in a number of processes involving DNA strand separation, including replication, repair, recombination, and transcription. Jarvis et al. (1996) identified a mouse gene encoding a novel putative helicase. They designated the gene LSH, for lymphoid-specific Hel, because Northern blot analysis revealed that it was expressed specifically in mouse early thymocytes. Geiman et al. (1998) determined that the mouse LSH, or HELLS (helicase lymphoid-specific), gene contains at least 18 exons and spans more than 26.6 kb. Using Northern blot analysis, these authors found that human HELLS is expressed in testis and thymus, two tissues that are highly active in DNA recombination. Geiman and Muegge (2000) demonstrated that LSH is not obligatory for normal lymphoid development but is essential for normal proliferation of peripheral T lymphocytes.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • 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

  • 一次抗体 備考

    DNA helicases (Hel) play a role in a number of processes involving DNA strand separation, including replication, repair, recombination, and transcription. Jarvis et al. (1996) identified a mouse gene encoding a novel putative helicase. They designated the gene LSH, for lymphoid-specific Hel, because Northern blot analysis revealed that it was expressed specifically in mouse early thymocytes. Geiman et al. (1998) determined that the mouse LSH, or HELLS (helicase lymphoid-specific), gene contains at least 18 exons and spans more than 26.6 kb. Using Northern blot analysis, these authors found that human HELLS is expressed in testis and thymus, two tissues that are highly active in DNA recombination. Geiman and Muegge (2000) demonstrated that LSH is not obligatory for normal lymphoid development but is essential for normal proliferation of peripheral T lymphocytes.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab3851 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
WB 1/2000 - 1/10000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).
IP Use a concentration of 2 - 4 µg/ml.

ターゲット情報

  • 機能

    Plays an essential role in normal development and survival. Involved in regulation of the expansion or survival of lymphoid cells. Required for de novo or maintenance DNA methylation. May control silencing of the imprinted CDKN1C gene through DNA methylation. May play a role in formation and organization of heterochromatin, implying a functional role in the regulation of transcription and mitosis.

  • 組織特異性

    Highly expressed in proliferative tissues such as adult thymus and testis, and expressed at lower levels in uterus, small intestine, colon, and peripheral blood mononuclear cells. Also expressed in neoplastic cell lines including those derived from myeloid and lymphoid leukemias.

  • 配列類似性

    Belongs to the SNF2/RAD54 helicase family.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.

  • 細胞内局在

    Nucleus. Closely associated with pericentric heterochromatin.
  • Information by UniProt

  • 参照データベース

    • 別名

      • FLJ10339 antibody
      • Helicase lymphoid specific antibody
      • Hells antibody
      • HELLS_HUMAN antibody
      • LSH antibody
      • Lymphoid specific helicase antibody
      • Lymphoid-specific helicase antibody
      • Nbla10143 antibody
      • PASG antibody
      • Proliferation associated SNF2 like protein antibody
      • Proliferation-associated SNF2-like protein antibody
      • SWI/SNF2 related matrix associated actin dependent regulator of chromatin subfamily A member 6 antibody
      • SWI/SNF2-related matrix-associated actin-dependent regulator of chromatin subfamily A member 6 antibody
      see all

    画像

    • Immunoprecipitation - Anti-SMARCA6 antibody (ab3851)
      Immunoprecipitation - Anti-SMARCA6 antibody (ab3851)

      Detection of Human HELLS by Western Blot of Immunoprecipitate using Affinity Purified Rabbit anti-HELLS (ab3851).

      Sample: Nuclear Extract (1mg) from HeLa cells

      Antibody: Affinity purified Rabbit anti-HELLS (ab3851)

      IP - 2µg. WB - 0.5µg/ml

      Detection: ECL, 30 seconds

       

    • Western blot - Anti-SMARCA6 antibody (ab3851)
      Western blot - Anti-SMARCA6 antibody (ab3851)
      Anti-SMARCA6 antibody (ab3851) at 1 µg/ml + Human thymus tissue lysate - total protein (ab30146) at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 97 kDa
      Observed band size: 97 kDa
      Additional bands at: 102 kDa, 120 kDa, 47 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 30 seconds

    参考文献 (1)

    ab3851 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。

    ab3851 は 1 報の論文で使用されています。

    • Jenness C  et al. HELLS and CDCA7 comprise a bipartite nucleosome remodeling complex defective in ICF syndrome. Proc Natl Acad Sci U S A 115:E876-E885 (2018). PubMed: 29339483

    レビューと Q&A

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    1-5 of 5 Abreviews or Q&A

    Question

    I was very concerned, but having received your message did spend several days of time and effort to test each of these antibodies. Unfortunately, none of the antibodies worked in IF. The high temperature and long transition time must have caused significant degradation. We order the vast majority of our antibodies from Abcam and have never had this problem before.

    I need your assistance in arranging for a replacement of these antibodies a soon as possible. I am under considerable time pressure to complete a series of experiments involving these antibodies, which are critical for an important upcoming grant deadline. It was also a significant cost of about ˜$1,200.00

    Please let me know what I can do to initiate this process. Is it possible to have the replacement antibodies shipped out to the lab? This would be a tremendous help!

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    Answer

    Thank you for confirming these details. I am sorry to hear that these antibodies did not work in ICC/IF.
    As requested, I have issued a free of charge replacement of ab3851, ab52060, and ab112511. These will be shipped out tonight for delivery tomorrow morning. It looks like that that original order was shipped on a Saturday for Monday's delivery. If you would prefer for us not to send orders on the weekend in the future, please let us know and we can make a note on your account.
    To check the status of the order please contact our Customer Service team and reference this number.
    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
    I wish you the best of luck with your research.

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    Western blot abreview for Anti-SMARCA6 antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MCF7 and HEK)
    Loading amount
    30 µg
    Specification
    MCF7 and HEK
    Gel Running Conditions
    Reduced Denaturing (8-16%)
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    投稿 Aug 02 2010

    Western blot abreview for Anti-SMARCA6 antibody

     Good
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Oocytes)
    Loading amount
    100 cells
    Specification
    Oocytes
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    投稿 Aug 30 2007

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SMARCA6 antibody

     Good
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Spermatogenic cells)
    Specification
    Spermatogenic cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.15% Triton X 100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Aug 29 2007

    Question

    Here is a complaint about ab3851/Anti-SMARCA6 with lot No 74811. Following is the detail. 1. Please describe the problem at first(when received the antibody) can get result with high background; but when stored it for 2 month, can’t get any bands. 2. On what material are you testing the antibody in WB? Species: human diploid fibroblast(young,middle,old) Cell extract or Nuclear extract: total Cell extract 3. The lysate How much protein was loaded: 120ug What lysis buffer was used: 50mM Tris-HCl,pH7.6?250mM NaCl? 5mM EDTA50mM NaF with protease inhibitors before using What protease inhibitors were used: 1% 100mM PMSF, appropriate leupeptin and aprotinin,10% NP-40 What loading buffer was used: 2×SDS loading buffer add beta-Me before using Did you heat the samples: temperature and time: 90-100C for 3-5mins 4. Electrophoresis/Gel conditions/ Transfer conditions Reducing or non reducing gel: Reducing Gel Gel percentage : 4% upper SDS-PAGE and 7.5% down SDS-PAGE Transfer conditions: Use Bio-Rad Trans-Blot Cell; Transfer is done at 250mA for 1.5hours on ice.; Transfer buffer----------Glycine 14.4g; methanol 100-200ml; Tris-base 3.03g; ddH2O up to 1000ml,keep in 4C 5. Blocking conditions Buffer: TBS-T(0.02% Tween-20) Blocking agent: milk, BSA, serum, what percentage: 5% milk Incubation time: 2hours Incubation temperature: RT 6. Primary Antibody Specification (in which species was it raised against): rabbit At what dilution(s) have you tested this antibody: 1:2000, 1:2500, 1:5000, 1:8000 What dilution buffer was used: TBS-T containing 1% no-fat milk Incubation time: overnight Incubation temperature: 4C What washing steps were done: 1×TBS-T for first time, in RT for 5min,then blocking buffer for 15min followed by 4 times wash with 1×TBS-T, each time 5min. All the times with shaking. 7. Secondary Antibody Specification (in which species was it raised against)? Goat anti rabbit IgG, HRP At what dilution(s) have you tested this antibody: 1:2000-1:5000 Incubation time: 4C for 1hours Wash steps: 1×TBS-T for first time, in RT for 5min,then blocking buffer for 15min followed by 4 times wash with 1×TBS-T, each time 5min. All the times with shaking Do you know whether the problems you are experiencing come from the secondary?: not from the secondary antibody for it worked well with other primary antibody 8. Detection method ECL 9. Background bands Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) yes 11. Did you apply positive and negative controls along with the samples? With primary antibody against b-actin, can get positive result. Using non-relative protein as negative control, get no bands Attchement is the result of before, with high background: from left to right: extract from Young 2BS,Middle 2BS, Old 2BS, Non-realtive protein

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    Answer

    I'm sorry to hear you are having a problem with ab3851. Thank you for taking the time to answer our questions and sending us an image of your blot, it is very helpful. I suspect the high background you had in the first blot is due to the presence of milk in the blocking step as well as in the dilution and washing buffers. I would recommend trying 5%BSA blocking and incubating the antibodies in TBST only. I would recommend testing a positive control of nuclear extract of Hela cell lysate to make sure levels of SMARCA6 in your samples are not below detection levels and to use ECL+ as this is more sensitive. The fact that the antibody did not work the second time after 2 months is very concerning, was the antibody stored at 4C and did it stay at room temperature for any significant amount of time? If you still have problems with these recommendations and with the positive control I can certainly offer you a replacement vial if you have purchased the antibody in the last 90 days,

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