Validated using a knockout cell line
Recombinant
RabMAb

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

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 within Mouse RING2/ RING1B/ RNF2 aa 150-250 (phospho S168). The exact sequence is proprietary.
    Database link: Q9CQJ4

  • 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

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

    This product is a recombinant rabbit monoclonal antibody.

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

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20902-58
  • Isotype

    IgG
  • Research areas

Applications

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.
  • 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.
    • Information by UniProt
    • Database links

    • 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

    • 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 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.

    • 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.

    • 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.

    References

    ab234421 has not yet been referenced specifically in any publications.

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