Anti-CLLD8/SETDB2抗体(ab5517)

Submit a review Q&A (2)References (3)

Key features and details

  • Rabbit polyclonal to CLLD8/SETDB2
  • Suitable for: ICC/IF
  • Reacts with: Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-CLLD8/SETDB2抗体
    参阅全部 CLLD8/SETDB2 一抗

  • 描述

    兔多克隆抗体to CLLD8/SETDB2

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IFmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide corresponding to Human CLLD8/SETDB2 aa 550-650 conjugated to keyhole limpet haemocyanin.
    Database link: Q96T68
    (Peptide available as ab24398)

  • 常规说明

    Our collaborator on this project has found that the antibody works poorly in WB.

性能

  • 形式

    Liquid

  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • 存储溶液

    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...

  • 纯度

    Immunogen affinity purified

  • 克隆

    多克隆

  • 同种型

    IgG

  • 研究领域

应用

Our Abpromise guarantee covers the use of ab5517 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF Use at an assay dependent concentration.

靶标

  • 功能

    Histone methyltransferase involved in left-right axis specification in early development and mitosis. Specifically trimethylates 'Lys-9' of histone H3 (H3K9me3). H3K9me3 is a specific tag for epigenetic transcriptional repression that recruits HP1 (CBX1, CBX3 and/or CBX5) proteins to methylated histones. Contributes to H3K9me3 in both the interspersed repetitive elements and centromere-associated repeats. Plays a role in chromosome condensation and segregation during mitosis.

  • 组织特异性

    Ubiquitous. Highest expression in heart, testis and ovary.

  • 序列相似性

    Belongs to the histone-lysine methyltransferase family.
    Contains 1 MBD (methyl-CpG-binding) domain.
    Contains 1 pre-SET domain.
    Contains 1 SET domain.

  • 细胞定位

    Nucleus. Chromosome.
  • Information by UniProt

  • 数据库链接

    • 别名

      • C13orf4 antibody
      • Chromosome 13 open reading frame 4 antibody
      • Chronic lymphocytic leukemia deletion region 8 antibody
      • Chronic lymphocytic leukemia deletion region gene 8 antibody
      • Chronic lymphocytic leukemia deletion region gene 8 protein antibody
      • Clld8 antibody
      • CLLL8 antibody
      • Gm293 antibody
      • H3-K9-HMTase antibody
      • Histone H3-K9 methyltransferase antibody
      • Histone-lysine N-methyltransferase SETDB2 antibody
      • KMT1F antibody
      • Lysine N-methyltransferase 1F antibody
      • Probable histone-lysine N-methyltransferase H3 lysine-9 specific antibody
      • SEB2 antibody
      • SET domain bifurcated 2 antibody
      • SET domain protein, bifurcated, 2 antibody
      • SETB2_HUMAN antibody
      • Setdb2 antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)
      Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)

      ICC/IF image of ab5517 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab5517 at 5ug/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti Rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)
      Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)This image is courtesy of Genevieve Fourel, Grenoble

      Left: Endogenous CLLD8

      Detection using indirect fluorescence of the signal corresponding to endogenous CLLD8 in 293T cells. Cells fixed using 4% formaldehyde, blocked with PBS containing 3% milk and 0.5% Triton X-100, incubated for 1 hour at 37 °C using a 1/50 dilution of antibody ab5517. Cells were then washed 3 times and incubated for 1 hour at 37 °C with a goat anti-rabbit secondary antibody (1/500 dilution) coupled to Alexa Fluor 555 (Molecular Probes). Following 3 more washes, cells were stained with DAPI and mounted with Vectashield.

      Top: DAPI
      Bottom: ab5517


      Right: Overexpressed CLLD8

      Detection using indirect fluorescence of the signal corresponding to staining with anti-FLAG mouse antibody (top) and an antibody (ab5517) against CLLD8 (middle) in 293T cells. 293T cells were transfected with vectors for overexpression of flagged CLLD8 using Lipofectamine 2000 transfection procedure (Invitrogen). Cells were fixed 48 hours post-tra

    • Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)
      Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)This image is courtesy of Genevieve Fourel, Grenoble

      Detection using indirect fluorescence of the signal corresponding to endogenous CLLD8 in 293T cells. Using Lipofectamine 2000 (Invitrogen), cells were transfected with either a control shRNA directed against luciferase (left, SiRluc) or a specific siRNA directed against CLLD8 (right). 48 hours post-transfection, cells were fixed, blocked, and incubated with a 1:50 dilution of ab5517 at 37 °C for 1 hour. Cells were then washed 3 times and incubated at 37 °C for 1 hour with a 1/500 dilution of goat anti-rabbit antibody coupled with Alexa Fluor 555 (Molecular Probes). Following 3 further washes cells were stained with DAPI and mounted with Vectashield.

      Antibody signals were extinguished when a specific RNAi was used. The few cells which retained a signal were presumably not transfected.   

       

    文献 (3)

    发表研究结果有使用 ab5517？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab5517 被引用在 3 文献中.

    • Nishikawaji T  et al. Oncogenic roles of the SETDB2 histone methyltransferase in gastric cancer. Oncotarget 7:67251-67265 (2016). IHC . PubMed: 27572307
    • Hogarth CA  et al. Identification and expression of potential regulators of the mammalian mitotic-to-meiotic transition. Biol Reprod 84:34-42 (2011). WB, IHC-P ; Mouse . PubMed: 20826732
    • Falandry C  et al. CLLD8/KMT1F is a lysine methyltransferase that is important for chromosome segregation. J Biol Chem 285:20234-41 (2010). WB, ICC/IF ; Human . PubMed: 20404330

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    Question

    I would like to request information on one of your antibodies, Anti-CLLD8/SETDB2 antibody (ab5517). I was wondering if this antibody works for mouse SETDB2

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    Answer

    Thank you for your inquiry. I BLASTed the immunogen sequence of ab5517 against mouse and it only has 55% homology.  We generally say that anything with a greater than 85% homology would be expected to cross-react, so this is quite low.  This antibody probably wouldn't work with mouse samples. However, we do have a goat polyclonal anti-CLLD8/SETDB2 antibody that is predicted to cross-react with mouse, ab13712.  This immunogen is 100% homologous with mouse. However, since we have not tested ab13712 in mouse, we would not be able to guarantee that it would work. https://www.abcam.com/CLLD8-SETDB2-antibody-ab13712.html But, I can offer a discount off a future purchase if you buy ab13712 now, test it in mouse and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab13712 in mouse.  I will then send a discount code. This code must be issued before purchasing ab13712 so please wait for my reply before ordering. 2. Purchase ab13712 either by phone, fax, or online (www.abcam.com). 3. Test it in mouse. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab13712 turns out to be unsuitable for mouse, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.  

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    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 72248 DESCRIPTION OF THE PROBLEM Also purchased ab13712. Multiple bands with each antibody. Molecular weight of bands different for both antibodies. Not sure if I'm picking up the correct band and if so with which antibody SAMPLE Human cell lysates A549, jurkat, C8166, Daudi, HUT-78 PRIMARY ANTIBODY ab5517 or ab13712 SECONDARY ANTIBODY cell signalling anti-Rabbit-HRP #7074 abcam rabbit anti-goat6741-1 DETECTION METHOD chemiglow POSITIVE AND NEGATIVE CONTROLS USED none available SAMPLE PREPARATION RIPA + protease inhibitors, heated 5min TRANSFER AND BLOCKING CONDITIONS NuPAGE blotting buffer + 20% MeOH, 2hr. Blocked 5% milk in PBS-tween HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No WHAT STEPS HAVE YOU ALTERED? primary antibody concentration

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    Answer

    I'm very sorry to hear you are having problems with ab5517 and ab13712. You do not mention adding loading buffer to your samples prior to boiling, please make sure you add loading buffer containing SDS, DTT, beta-mercaptoethanol and bromphenol blue. Please also make sure you keep samples on ice until boiling and that protease inhibitors are added freshly. You do not mention how long you block the membrane for (and what temperature), I recommend trying 5%BSA in TBST for 1 hr, then rinsing a few seconds in TBST and incubating both primary and secondary antibodies in TBST. The dilution recommended for ab5517 is 1:5000 and for ab13712 is ug/ml but you may need to dilute more, I hope these suggestions help, please do not hesitate to contact us if you need further assistance,

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