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  1. Link

    ring2--ring1b--rnf2-phospho-s168-antibody-epr20902-58-ab234421.pdf

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Epigenetics and Nuclear Signaling Chromatin Remodeling Polycomb Silencing PRC1
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)

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  • SDS
  • Certificate of Compliance
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Western blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
  • Dot Blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
  • Immunoprecipitation - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
  • Western blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
  • Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20902-58] to RING2 / RING1B / RNF2 (phospho S168)
  • Suitable for: WB, IP, Dot blot
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Knockout
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Human RNF2 (RING2 / RING1B) knockout HeLa cell line (ab264845)

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Overview

  • Product name

    Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58]
    See all RING2 / RING1B / RNF2 primary antibodies
  • Description

    Rabbit monoclonal [EPR20902-58] to RING2 / RING1B / RNF2 (phospho S168)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Dot blotmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Overexpression of wild type RNF2 in RNF2 knock out mouse embryonic stem cells derived from C57BL/6 mouse, whole cell lysate; NCCIT, HEK-293T, HepG2 and F9 whole cell lysates; mouse testis tissue lysate. Dot blot: RNF2 (phospho S168) peptide (aa161-172) and RNF2 (phospho S168) peptide (aa165-176). IP: F9 whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20902-58
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] - BSA and Azide free (ab256315)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

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

    • Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (ab181140)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab234421 in the following tested applications.

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

Application Abreviews Notes
WB
1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 38 kDa).
IP
1/30.
Dot blot
1/1000.
Notes
WB
1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 38 kDa).
IP
1/30.
Dot blot
1/1000.
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

Target

  • Function

    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.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Contains 1 RING-type zinc finger.
  • Post-translational
    modifications

    Polyubiquitinated in the presence of UBE2D3 (in vitro).
    Monoubiquitinated, by auto-ubiquitination.
  • Cellular localization

    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
  • Database links

    • Entrez Gene: 6045 Human
    • Entrez Gene: 19821 Mouse
    • Omim: 608985 Human
    • SwissProt: Q99496 Human
    • SwissProt: Q9CQJ4 Mouse
    • Unigene: 591490 Human
    • Unigene: 482250 Mouse
    • Alternative names

      • 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

    Images

    • Western blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      Western blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      All lanes : Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421) at 1/1000 dilution

      Lane 1 : Overexpression of wild type RNF2 in RNF2 knock out mouse embryonic stem cells derived from C57BL/6 mouse, whole cell lysate
      Lane 2 : Overexpression of S168A mutant RNF2 in RNF2 knock out mouse embryonic stem cells derived from C57BL/6 mouse, whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 38 kDa
      Observed band size: 40 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer:  5% NFDM/TBST.

      Lysates were kindly provided by Dr Jinzhong Qin.

    • Dot Blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      Dot Blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)

      Dot blot analysis of RING2 / RING1B / RNF2 (phospho S168) peptide labeled with ab234421 at 1/1000 dilution.

      Lane 1: RING2 / RING1B / RNF2 (phospho S168) peptide (aa161-172).
      Lane 2: RING2 / RING1B / RNF2 (phospho S168) peptide (aa165-176).
      Lane 3: RING2 / RING1B / RNF2 non-phospho peptide (aa161-172).

      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Immunoprecipitation - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      Immunoprecipitation - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)

      RING2 / RING1B / RNF2 (phospho S168) was immunoprecipitated from 0.35 mg of F9 (mouse embryonic testicular cancer cell line) whole cell lysate with a234421 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab234421 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

      Lane 1: F9 whole cell lysate 10 μg (Input).
      Lane 2: ab234421 IP in F9 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab234421 in F9 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 30 seconds.

    • Western blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      Western blot - Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      All lanes : Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421) at 1/1000 dilution

      Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : NCCIT (human pluripotent embryonic carcinoma cell line) whole cell lysate
      Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 5 : F9 (mouse embryonic testicular cancer cell line) whole cell lysate
      Lane 6 : Mouse testis tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 38 kDa
      Observed band size: 40 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lane 1: 3 minutes; Lanes 2 & 3: 15 seconds; Lanes 4-6: 3 minutes.

    • Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)
      Anti-RING2 / RING1B / RNF2 (phospho S168) antibody [EPR20902-58] (ab234421)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab234421? Please let us know so that we can cite the reference in this datasheet.

    ab234421 has not yet been referenced specifically in any publications.

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