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使用敲除细胞株进行验证

Anti-CBX1 / HP1 beta抗体[MAC353] (ab10811)

Reviews (2)Q&A (3)References (15)
Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)

Key features and details

  • Rat monoclonal [MAC353] to CBX1 / HP1 beta
  • Suitable for: WB, ICC/IF, IP, ELISA, IHC-Fr, ChIP
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG2b

概述

  • 产品名称

    Anti-CBX1 / HP1 beta抗体[MAC353]
    参阅全部 CBX1 / HP1 beta 一抗

  • 描述

    大鼠单克隆抗体[MAC353] to CBX1 / HP1 beta

  • 宿主

    Rat

  • 特异性

    Ab10811 recognises the M31 molecule in mouse and the homologous HP1 HS beta molecule in man.

  • 经测试应用

    适用于: WB, ICC/IF, IP, ELISA, IHC-Fr, ChIPmore details

  • 种属反应性

    与反应: Mouse, Human

  • 免疫原

    Fusion protein corresponding to Mouse CBX1/ HP1 beta (C terminal).
    Database link: P83917

  • 阳性对照

    • Murine and human nuclear extracts.

  • 常规说明

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab10811于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (1)
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 22.2 kDa).
ICC/IF (1)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
说明
WB
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 22.2 kDa).
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.

靶标

  • 功能

    Component of heterochromatin. Recognizes and binds histone H3 tails methylated at 'Lys-9', leading to epigenetic repression. Interaction with lamin B receptor (LBR) can contribute to the association of the heterochromatin with the inner nuclear membrane.

  • 组织特异性

    Expressed in all adult and embryonic tissues.

  • 序列相似性

    Contains 2 chromo domains.

  • 翻译后修饰

    Not phosphorylated.
    Ubiquitinated.

  • 细胞定位

    Nucleus. Unassociated with chromosomes during mitosis.
  • Information by UniProt

  • 数据库链接

    • 别名

      • CBX 1 antibody
      • CBX antibody
      • Cbx1 antibody
      • CBX1_HUMAN antibody
      • Chromobox 1 antibody
      • Chromobox homolog 1 (HP1 beta homolog Drosophila ) antibody
      • chromobox homolog 1 antibody
      • Chromobox protein homolog 1 antibody
      • Drosophila HP1 beta antibody
      • Heterochromatin protein 1 beta antibody
      • Heterochromatin protein 1 homolog beta antibody
      • Heterochromatin protein p25 antibody
      • heterochromatin protein p25 beta antibody
      • HP1 beta antibody
      • HP1 beta homolog antibody
      • HP1 beta homolog Drosophila antibody
      • HP1, Drosophila. homolog of, beta antibody
      • HP1beta antibody
      • HP1Hs beta antibody
      • HP1Hsbeta antibody
      • Human heterochromatin protein p25 mRNA complete cds antibody
      • M31 antibody
      • MOD 1 antibody
      • MOD1 antibody
      • Modifier 1 protein antibody
      • p25beta antibody
      see all

    图片

    • Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
      Western blot - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)

      Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: CBX1 knockout HAP1 cell lysate (40 µg)
      Lane 3: MCF7 cell lysate (40 µg)
      Lane 4: A431 cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab10811 observed at 26 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab10811 was shown to specifically react with CBX1 / HP1 beta when CBX1 / HP1 beta knockout samples were used. Wild-type and CBX1 / HP1 beta knockout samples were subjected to SDS-PAGE. Ab10811 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rat IgG H&L (IRDye® 800CW) (ab253031) preadsorbed and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)
      Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)This image is courtesy of Scott Slattery and Mike Mancini
      HeLA cells were stained with ab10811 in panel one. In panel two they were stained with ab10811 (green), DAPI (blue) and SH-CREST (red), which stains the centromeres. Fix 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC diluted 1/100 in 5% milk in TBST. Secondary antibody incubated 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
    • Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)
      Immunocytochemistry/ Immunofluorescence - Anti-CBX1 / HP1 beta antibody [MAC353] (ab10811)Image from Talasz H et al., J Biol Chem. 2005 Nov 18;280(46):38814-22. Epub 2005 Sep 15. Fig 1.; doi: 10.1074/jbc.M505563200; November 18, 2005 The Journal of Biological Chemistry, 280, 38814-38822.
      Immunofluorescence analysis of mouse erythroleukemia cell nuclei, staining CBX1 / HP1 beta in heterochromatin, with ab10811.

      Cells were fixed with paraformaldehyde, permeabilized using Triton X-100 and blocked for 30 min with 2.5% BSA. Cells were incubated with primary antibody (1/50) before incubating with a Cy3-conjugated goat anti-rat IgG to detect staining.

    文献 (15)

    发表研究结果有使用 ab10811？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab10811 被引用在 15 文献中.

    • Lamb KN  et al. Discovery of Potent Peptidomimetic Antagonists for Heterochromatin Protein 1 Family Proteins. ACS Omega 7:716-732 (2022). PubMed: 35036738
    • Qin W  et al. Probing protein ubiquitination in live cells. Nucleic Acids Res 50:e125 (2022). PubMed: 36189882
    • Waybright JM  et al. A Peptidomimetic Ligand Targeting the Chromodomain of MPP8 Reveals HRP2's Association with the HUSH Complex. ACS Chem Biol 16:1721-1736 (2021). PubMed: 34415726
    • Otake K  et al. CENP-B creates alternative epigenetic chromatin states permissive for CENP-A or heterochromatin assembly. J Cell Sci 133:N/A (2020). PubMed: 32661090
    • Yang G  et al. The histone H3K9 methyltransferase SUV39H links SIRT1 repression to myocardial infarction. Nat Commun 8:14941 (2017). PubMed: 28361889
    View all Publications for this product

    客户评价及客户问答

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    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (RAW264.7 cells)
    Gel Running Conditions
    Reduced Denaturing (14 %)
    Loading amount
    80 µg
    Specification
    RAW264.7 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Magali Boissiere

    Verified customer

    提交于 Feb 22 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade

     Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Colorectal Cancer Cells)
    Specification
    Colorectal Cancer Cells
    Fixative
    Formaldehyde
    Permeabilization
    Yes - triton x-100
    Blocking step
    Serum as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Jan 21 2010

    Question

    Hi，
    I ordered a primary antibody (Anti-CBX1 / HP1 beta antibody [MAC353] - ChIP Grade (ab10811)) and I plan to use this Ab for ChIP. But the concentration of this Ab is unknown. I wanna ask whether you have any reference data for ChIP using this Ab.
    Thank you very much!


    --

    Read More
    Answer

    Thank you for contacting us.


    For ChIP, we recommend to optimise the working concentration according to sample and experiment. At the starting point, you could try using 1:50 dilution.


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Question

    I just wanted to know if this antibody CBX1 MAC 353 (ab10811) is only detecting CBX1 without phosphorylated sites on Thr51 or with phosphorylated sites on Thr51? Or is it just the whole amount of CBX1-protein?

    Read More
    Answer

    Thank you for contacting us with your questions. Since the immunogen of the antibody was taken from the C-terminus, and threonine 51 is much closer to the N-terminus, the antibody will detect the total CBX1 protein. The antibody will recognize CBX1 whether or not it is phosphorylated at threonine 51. I hope this information is helpful, but please let me know if you have any further questions.

    Read More

    Question

    I received antibody, and thanks for the pretty mug cup. I like to know the concentration.What's the recommended dilution for western and immunofluorescence?

    Read More
    Answer

    I'm glad you like the mug! We are unable to provide a concentration for ab10811 as it is in an unpurified form, tissue culture supernatant, and we cannot guarantee the concentration for unpurified material. I recommend starting at a dilution of 1:1000 and optimizing from there.

    Read More

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