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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ubiquitin E3 Enzymes RING Finger E3 Ligase

Anti-Mel18抗体(ab5267)

  • Datasheet
Reviews (3)Q&A (1)References (14)

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Western blot - Anti-Mel18 antibody (ab5267)

    Key features and details

    • Goat polyclonal to Mel18
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

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    概述

    • 产品名称

      Anti-Mel18抗体
    • 描述

      山羊多克隆抗体to Mel18
    • 宿主

      Goat
    • 经测试应用

      适用于: WBmore details
    • 种属反应性

      与反应: Human
    • 免疫原

      Synthetic peptide corresponding to Human Mel18 aa 300-400 (C terminal).
      Database link: NP_009075.1

      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
    • 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

    • 形式

      Liquid
    • 存放说明

      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.
    • 存储溶液

      pH: 7.30
      Preservative: 0.02% Sodium azide
      Constituents: Tris buffered saline, 0.5% BSA
    • Concentration information loading...
    • 纯度

      Immunogen affinity purified
    • 纯化说明

      Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
    • 克隆

      多克隆
    • 同种型

      IgG
    • 研究领域

      • Cell Biology
      • Proteolysis / Ubiquitin
      • Proteasome / Ubiquitin
      • Ubiquitin E3 Enzymes
      • RING Finger E3 Ligase
      • Epigenetics and Nuclear Signaling
      • Chromatin Remodeling
      • Polycomb Silencing
      • PRC1
      • Epigenetics and Nuclear Signaling
      • Transcription
      • Cancer susceptibility
      • Tumor Suppressors
      • Stem Cells
      • Hematopoietic Progenitors
      • Surface Molecules

    相关产品

    • ChIP Related Products

      • ChIP Kit (ab500)
    • Compatible Secondaries

      • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
      • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
    • Isotype control

      • Goat IgG, polyclonal - Isotype Control (ab37373)

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab5267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
    Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).

    A 1 hour primary incubation is recommended for this product.

    说明
    WB
    Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).

    A 1 hour primary incubation is recommended for this product.

    靶标

    • 功能

      Transcriptional repressor. Binds specifically to the DNA sequence 5'-GACTNGACT-3'. Has tumor suppressor activity. May play a role in control of cell proliferation and/or neural cell development. Regulates proliferation of early T progenitor cells by maintaining expression of HES1. Also plays a role in antero-posterior specification of the axial skeleton and negative regulation of the self-renewal activity of hematopoietic stem cells. Component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility.
    • 组织特异性

      Detected in all tissues examined with high expression found in placenta lung and kidney and low expression, in liver, pancreas and skeletal muscle.
    • 序列相似性

      Contains 1 RING-type zinc finger.
    • 翻译后修饰

      Phosphorylated. Homodimer formation is regulated by phosphorylation state with unphosphorylated protein forming homodimers.
    • 细胞定位

      Nucleus.
    • Target information above from: UniProt accession P35227 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • 数据库链接

      • Entrez Gene: 7703 Human
      • Omim: 600346 Human
      • SwissProt: P35227 Human
      • Unigene: 371617 Human
      • Unigene: 713595 Human
      • 别名

        • DNA binding protein Mel 18 antibody
        • DNA-binding protein Mel-18 antibody
        • Mel 18 antibody
        • MGC10545 antibody
        • PCGF 2 antibody
        • PCGF2 antibody
        • PCGF2_HUMAN antibody
        • Polycomb group ring finger 2 antibody
        • Polycomb group RING finger protein 2 antibody
        • RING finger protein 110 antibody
        • RNF110 antibody
        • Zinc finger protein 144 antibody
        • ZNF 144 antibody
        • ZNF144 antibody
        see all

      图片

      • Western blot - Anti-Mel18 antibody (ab5267)
        Western blot - Anti-Mel18 antibody (ab5267)

        ab5267 staining (1µg/ml) of Human Lung lysate (RIPA buffer, 30µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence.

        ab5267 staining (1µg/ml) of Human Lung lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.

      实验方案

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

      Click here to view the general protocols

      数据表及文件

      • Datasheet download

        Download

      文献 (14)

      发表研究结果有使用 ab5267?请让我们知道,以便我们可以引用本数据表中的参考文章。

      ab5267 被引用在 14 文献中.

      • Paschos K  et al. Requirement for PRC1 subunit BMI1 in host gene activation by Epstein-Barr virus protein EBNA3C. Nucleic Acids Res N/A:N/A (2019). PubMed: 30649516
      • Wang J  et al. Co-inhibition of BMI1 and Mel18 enhances chemosensitivity of esophageal squamous cell carcinoma in vitro and in vivo. Oncol Lett 17:5012-5022 (2019). PubMed: 31186712
      • Klauke K  et al. Polycomb Cbx family members mediate the balance between haematopoietic stem cell self-renewal and differentiation. Nat Cell Biol 15:353-62 (2013). WB . PubMed: 23502315
      • Holdt LM  et al. Alu elements in ANRIL non-coding RNA at chromosome 9p21 modulate atherogenic cell functions through trans-regulation of gene networks. PLoS Genet 9:e1003588 (2013). PubMed: 23861667
      • Lavial F  et al. Bmi1 facilitates primitive endoderm formation by stabilizing Gata6 during early mouse development. Genes Dev 26:1445-58 (2012). Mouse . PubMed: 22713603
      View all Publications for this product

      客户评价及客户问答

      Show All 评价 Q&A
      提交评价 提交问题

      1-4 of 4 Abreviews or Q&A

      ChIP abreview for Anti-Mel18 antibody - ChIP Grade

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      ChIP
      Sample
      Human Cell lysate - whole cell (NT2 cell line)
      Negative control
      Intergenic region on Chromosome 11.
      Specification
      NT2 cell line
      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: 1% Formaldehyde
      Positive control
      CCND2 and ALX4 genes
      Read More

      Tatiana Kan

      Verified customer

      提交于 Oct 17 2016

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Mel18 antibody

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (NB4 cell line(acute promylocytic leukemia))
      Specification
      NB4 cell line(acute promylocytic leukemia)
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - 0.1% Triton X-100
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
      Read More

      Mr. Sungsin Jo

      Verified customer

      提交于 Oct 03 2012

      ChIP abreview for Anti-Mel18 antibody

      Good
      Abreviews
      Abreviews
      Application
      ChIP
      Sample
      Human Cell lysate - whole cell (HL60(acute promyelotic leuemia cells line))
      Specification
      HL60(acute promyelotic leuemia cells line)
      Type
      Cross-linking (X-ChIP)
      Duration of cross-linking step: 10 minute(s) and 0 second(s)
      Specification of the cross-linking agent: SDS lysis buffer
      Detection step
      Semiquantitative PCR
      Positive control
      acetly-histone H3 (cat #06-599, millipore)
      Negative control
      Rabbit IgG(cat# pp64, millipore)
      Read More

      Mr. Sungsin Jo

      Verified customer

      提交于 Oct 23 2009

      Question

      A few weeks ago I ordered two antibodies anti-Mel18 (ab5267) and anti-Ring1B (ab3832). I have tried using them on HeLa nuclear extract and HeLa chromatin but I am not getting any signal. therefore, I have few questions regarding those antibodies:1 What positive controls do you use to find out if those antibodies are working? 2.What western blot conditions do you use? (antibody dilutions,membrane type, blotting conditions) I am also wondering if you could send me one of your positive controls so I could detect the proteins using the above mentioned antibodies. also, if you have another lot of these antibodies available I would appreciate getting them as well. I have very small amounts of tumor samples that I need to check for Mel18 and Ring1B levels and I want to make sure the antibodies are working before I do that.

      Read More

      Abcam community

      Verified customer

      Asked on Oct 11 2005

      Answer

      Thank you for your enquiry. I have consulted the EXPASY databases and I retrieved the following information: Mel18 - Tissue specificity: Detected in all tissues examined with high expression found in placenta, lung and kidney and low expression, in liver, pancreas and skeletal muscle. Ring1B - No tissue specificity information. The recommended positive control for Mel18 is human lung lysate. For Ring1B we do not have a recommended positive control. However, a reviewer has successfully used this antiserum in western blotting using mouse cultured cells (3T3) and mouse brain. This may be a useful alternative to HeLa cells as I cannot guarantee that these cells express Ring1B. I have obtained the following details of how the positive control to how Mel18 was prepared and run by western blotting. Western Blot: Approx 40 kDa band observed in Human Lung extracts (predicted MW of 40kDa according to NP_009075). Recommended for use at 1-2µg/ml. Tissue Lysis. Tissue chunks were weighed and cut into approx 1mm cubes using a razor blade. The tissue was transferred to a handheld homogenizer and 3 ml of ice-cold RIPA buffer was added per 1g of tissue. The tissue was gently homogenised over 20 minutes on ice. The resulting lysate was aliquotted into 1.5 ml microfuge tubes and centrifuged at 13,000 rpm for 5 min in a microfuge. The supernatant was transferred into clean tubes and its protein concentration was measured with BioRad protein assay. The concentration was then adjusted to 5 mg/ml with RIPA lysis buffer. An equal volume of 2 x SDS sample buffer was added and the cell lysate was boiled for 5 minutes. Lysates were stored at -80C until use.(RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton X-100, 1% Sodium deoxycholate, 0.1% SDS). SDS PAGE. Samples were run at 200V constant on a 12% acrylamide SDS-PAGE mini gel - using Biorad Mini-Protean 3 kit and protocols. Before loading samples had 5% (v/v) 2-ME added and were boiled for 3 minutes. Transfer. We used a Biorad Mini Trans-Blot, constant 100 V for 1 hour. Transfer Buffer was 20 mM Tris pH 8.0, 150 mM Glycine, 10% Methanol. We transferred to Millipore PVDF membrane and stained with Ponceau Red to evaluate the transfer. Staining. The membrane was blocked in 2.5% skimmed milk in TBS-T (TBS + 0.05% Tween-20) for 1 hr at room temperature with agitation. Primary antibody was incubated for 1 hr at room temperature with agitation. We used [a competitor's] secondary 1:3000) for 1 hr at room temperature with agitation. We washed with TBST three times after primary and secondary antibody, each wash lasting for 5-10 mins. ECL-plus (Amersham) was used rather than ECL, which is considerably more sensitive. Final detection was on autoradiography film. I hope this information helps. Please do not hesitate to contact me should you require further assistance.

      Read More

      Abcam Scientific Support

      回复于 Oct 17 2005

      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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