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Epigenetics and Nuclear Signaling Chromatin Remodeling Polycomb Silencing PRC1

Anti-RING2 / RING1B / RNF2抗体(ab3832)

  • Datasheet
  • SDS
Reviews (2)Q&A (3)References (23)

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Western blot - Anti-RING2 / RING1B / RNF2 antibody - ChIP Grade (ab3832)

    Key features and details

    • Goat polyclonal to RING2 / RING1B / RNF2
    • Suitable for: WB
    • Reacts with: Mouse, Human, Chinese hamster
    • Isotype: IgG

    选择批间可重复性更高的重组抗体

    Product image
    Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (ab181140)
    • 研究可靠 —— 各批次间结果一致且可重复
    • 长期批量供应 —— 采用重组技术,可实现快速生产
    • 首次实验即可成功 —— 经过大量验证确认了特异性
    • 符合伦理标准 —— 产品不含动物成分

    概述

    • 产品名称

      Anti-RING2 / RING1B / RNF2抗体
      参阅全部 RING2 / RING1B / RNF2 一抗
    • 描述

      山羊多克隆抗体to RING2 / RING1B / RNF2
    • 宿主

      Goat
    • 特异性

      This antibody recognises a band at just above the expected size of 37kD.
    • 经测试应用

      适用于: WBmore details
    • 种属反应性

      与反应: Mouse, Human, Chinese hamster
    • 免疫原

      Synthetic peptide:

      EAGPSNKRTKTSDC

      , corresponding to amino acids 189-201 of Human RING1B.
      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
    • 常规说明

      RING1B interacts with CP2, a mammalian member of the grainyhead-like family of transcription factors, in vitro and in vivo. The functional consequence of this interaction is repression of CP2-dependent transcription.

      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

      Preservative: 0.01% Sodium azide
      Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
    • Concentration information loading...
    • 纯度

      Immunogen affinity purified
    • Primary antibody说明

      RING1B interacts with CP2, a mammalian member of the grainyhead-like family of transcription factors, in vitro and in vivo. The functional consequence of this interaction is repression of CP2-dependent transcription.
    • 克隆

      多克隆
    • 同种型

      IgG
    • 研究领域

      • Epigenetics and Nuclear Signaling
      • Chromatin Remodeling
      • Polycomb Silencing
      • PRC1

    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)
    • Recombinant Protein

      • Recombinant Human RING2 / RING1B / RNF2 protein (ab159360)

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab3832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 43 kDa (predicted molecular weight: 37.6 kDa).
    说明
    WB
    Use at an assay dependent concentration. Detects a band of approximately 43 kDa (predicted molecular weight: 37.6 kDa).

    靶标

    • 功能

      E3 ubiquitin-protein ligase that mediates monoubiquitination of 'Lys-119' of histone H2A, thereby playing a central role in histone code and gene regulation. H2A 'Lys-119' ubiquitination gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. May be involved in the initiation of both imprinted and random X inactivation. Essential 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 act via chromatin remodeling and modification of histones, rendering chromatin heritably changed in its expressibility. E3 ubiquitin-protein ligase activity is enhanced by BMI1/PCGF4. Acts as the main E3 ubiquitin ligase on histone H2A of the PRC1 complex, while RING1 may rather act as a modulator of RNF2/RING2 activity.
    • 通路

      Protein modification; protein ubiquitination.
    • 序列相似性

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

      Polyubiquitinated in the presence of UBE2D3 (in vitro).
      Monoubiquitinated, by auto-ubiquitination.
    • 细胞定位

      Nucleus. Chromosome. Enriched on inactive X chromosome (Xi) in female trophoblast stem (TS) cells as well as differentiating embryonic stem (ES) cells. The enrichment on Xi is transient during TS and ES cell differentiation. The association with Xi is mitotically stable in non-differentiated TS cells.
    • Target information above from: UniProt accession Q99496 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: 6045 Human
      • Entrez Gene: 19821 Mouse
      • Omim: 608985 Human
      • SwissProt: Q99496 Human
      • SwissProt: Q9CQJ4 Mouse
      • Unigene: 591490 Human
      • Unigene: 482250 Mouse
      • 别名

        • BAP 1 antibody
        • BAP1 antibody
        • DING antibody
        • DinG protein antibody
        • E3 ubiquitin protein ligase RING 2 antibody
        • E3 ubiquitin protein ligase RING2 antibody
        • E3 ubiquitin-protein ligase RING2 antibody
        • HIP2 interacting protein 3 antibody
        • HIP2-interacting protein 3 antibody
        • HIPI 3 antibody
        • HIPI3 antibody
        • Huntingtin interacting protein 2 interacting protein 3 antibody
        • Huntingtin-interacting protein 2-interacting protein 3 antibody
        • OTTHUMP00000060668 antibody
        • Polycomb M33 interacting protein Ring 1B antibody
        • Polycomb M33 interacting protein Ring1B antibody
        • Protein DinG antibody
        • RING 1B antibody
        • RING 2 antibody
        • RING finger protein 1B antibody
        • RING finger protein 2 antibody
        • RING finger protein BAP 1 antibody
        • RING finger protein BAP-1 antibody
        • RING finger protein BAP1 antibody
        • RING1b antibody
        • RING2_HUMAN antibody
        • RNF 2 antibody
        • Rnf2 antibody
        see all

      图片

      • Western blot - Anti-RING2 / RING1B / RNF2 antibody - ChIP Grade (ab3832)
        Western blot - Anti-RING2 / RING1B / RNF2 antibody - ChIP Grade (ab3832)

        Western blot using ab3832 at 1/500. Lysates at 20µg/lane.

        Lane 1: 3T3 cell lysate
        Lane 2: U937 cell lysate
        Lane 3: Jurkat cell lysate
        Lane 4: Mouse brain lysate
        Lane 5: CHO-K1 cell lysate

        Secondary ab: rabbit polyclonal to goat IgG ab6741 1/5000
        Exposure time: 3 minutes

        Western blot using ab3832 at 1/500. Lysates at 20µg/lane.

        Lane 2: U937 cell lysate
        Lane 3: Jurkat cell lysate
        Lane 4: Mouse brain lysate
        Lane 5: CHO-K1 cell lysate

        Secondary ab: rabbit polyclonal to goat IgG ab6741 1/5000 Exposure time: 3 minutes

      实验方案

      • Western blot protocols

      Click here to view the general protocols

      数据表及文件

      • SDS download

      • Datasheet download

        Download

      文献 (23)

      发表研究结果有使用 ab3832?请让我们知道,以便我们可以引用本数据表中的参考文章。

      ab3832 被引用在 23 文献中.

      • Plys AJ  et al. Phase separation of Polycomb-repressive complex 1 is governed by a charged disordered region of CBX2. Genes Dev 33:799-813 (2019). PubMed: 31171700
      • Papp B  et al. Genome-wide identification of direct RTA targets reveals key host factors for KSHV lytic reactivation. J Virol N/A:N/A (2018). PubMed: 30541837
      • Rothberg JLM  et al. Mtf2-PRC2 control of canonical Wnt signaling is required for definitive erythropoiesis. Cell Discov 4:21 (2018). PubMed: 29736258
      • Pulikkan JA  et al. CBFß-SMMHC Inhibition Triggers Apoptosis by Disrupting MYC Chromatin Dynamics in Acute Myeloid Leukemia. Cell 174:172-186.e21 (2018). PubMed: 29958106
      • Kang H  et al. Bivalent complexes of PRC1 with orthologs of BRD4 and MOZ/MORF target developmental genes in Drosophila. Genes Dev 31:1988-2002 (2017). PubMed: 29070704
      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

      1-5 of 5 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-RING2 / RING1B / RNF2 antibody - ChIP Grade

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (Hela cells)
      Specification
      Hela cells
      Fixative
      Methanol
      Permeabilization
      No
      Blocking step
      Fish scale gelatin in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · 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.

      Abcam user community

      Verified customer

      提交于 Aug 02 2010

      Western blot abreview for Anti-RING2 / RING1B / RNF2 antibody - ChIP Grade

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (Jurkat cells)
      Loading amount
      100000 cells
      Specification
      Jurkat cells
      Gel Running Conditions
      Reduced Denaturing
      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.

      Abcam user community

      Verified customer

      提交于 May 17 2010

      Question

      dear abcam,

      in May I bought RING1b (ab3832) antibody and accordingly to the datasheet it should work also for westerblot.
      unfortunately, after trying twice i did not successes to have a band or a signal after HCL exposure.
      i loaded 20 ug of RIPA lysed cells and i diluted the antibody 1:500 in PBS-Tween for overnight incubation.
      under this regard, do you have any suggestion or really this antibody does not work for western blot?
      looking forward for a positive answer form you,
      best regards,

      Read More

      Abcam community

      Verified customer

      Asked on Sep 26 2012

      Answer

      Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

      I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and in human and mouse samples. In the event that a product is not functioning in the tested applications and species 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 would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records.with furhter information, I woudl be more able to provide further suggestion onthe western blot. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you are able to complete this. It will help you put the information we require together very easily.

      Thank you for your time and cooperation. We look forward to receiving the completed questionaire.



      Order Details
      Antibody code:RING1b (ab3832) antibody

      Problem
      Choose: Non-specific band Multiple bands No signal or weak signal High background

      Lot number

      Purchase order number
      or preferably Abcam order number:



      General Information
      Antibody storage conditions (temperature/reconstitution etc)


      Description of the problem (high background, wrong band size, more bands, no band etc.)


      Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


      Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


      Amount of protein loaded
      20µg

      Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


      Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


      Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

      1:500 in PBS-Tween for overnight incubation.

      Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


      Detection method (ECL, ECLPlus etc.)

      HCL exposure

      Positive and negative controls used (please specify)



      Optimization attempts (problem solving)
      How many times have you tried the Western?



      Have you run a "No Primary" control?
      Yes No

      Do you obtain the same results every time?
      Yes No
      e.g. are the background bands always in the same place?


      What steps have you altered?


      Additional Notes:


      Image:
      We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

      Read More

      Abcam Scientific Support

      回复于 Sep 26 2012

      Question

      Inquiry: I am trying to ChIP Ring1b from mouse ES cells. The cells are crossliked with 1% formaldehyde, quenched with glycine (0.14M) and sonicated using a misonix sonicator. DNA fragments following sonication are on average 250 bp. I am using 20milliion ES cells per IP, and using massive amounts (up to 40ug per IP) of your product ab3832 to IP. However, I cannot detect the presence of the protein IP'ed even when I load the entire IP sample. The anitbody is bound to the antigen in the lysate, then protein G dynabeads are used to capture the Ab-Ag complexes. I fail to detect an IP product even when I perform no washes. Is there another Ab that actually works for ChIP? I have also tried using lot GR86503-1, with no IP as well.

      Read More

      Abcam community

      Verified customer

      Asked on Jun 14 2012

      Answer

      Thank you for contacting us.
      I am sorry to hear the antibody does not give you results as expected.

      I do have a couple questions in order to understand your experiment better:

      1) Are you using the antibody for IP or ChIP? There are differences between both techniques including the cross-linking step and the detection method. IP by itself has not been tested with this antibody.

      2) Are you using WB (for IP) to obtain your results or rather RT-PCR (for ChIP)?
      3) Also, what controls have you included in your experiment, especially when doing ChIP?
      4) Could you please send me your protocol?

      I look forward to hear back from you.

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

      Read More

      Abcam Scientific Support

      回复于 Jun 14 2012

      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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