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    ring1-antibody-epr13047-ab180170.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-RING1 antibody [EPR13047] (ab180170)

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Immunoprecipitation - Anti-RING1 antibody [EPR13047] (ab180170)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] (ab180170)
  • Western blot - Anti-RING1 antibody [EPR13047] (ab180170)
  • Western blot - Anti-RING1 antibody [EPR13047] (ab180170)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] (ab180170)
  • Immunocytochemistry/ Immunofluorescence - Anti-RING1 antibody [EPR13047] (ab180170)
  • Anti-RING1 antibody [EPR13047] (ab180170)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13047] to RING1
  • Suitable for: IHC-P, WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-RING1 antibody [EPR13047]
    See all RING1 primary antibodies
  • Description

    Rabbit monoclonal [EPR13047] to RING1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Dog, Chimpanzee, Rhesus monkey, Gorilla
  • Immunogen

    Synthetic peptide within Human RING1 aa 150-250 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q06587

  • Positive control

    • Human endometrial carcinoma and pancreas tissues. Ramos, MOLT4, LNCaP and HeLa whole cell lysate (ab150035). HeLa cells.
  • General notes

    Mouse and Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR13047
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-RING1 antibody [EPR13047] - BSA and Azide free (ab250174)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Ramos whole cell lysate (ab3955)
    • MOLT-4 whole cell lysate (ab7912)
  • Recombinant Protein

    • Recombinant Human RING1 protein (ab112348)
  • Related Products

    • Recombinant Human RING1 protein (ab112348)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab180170 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
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Predicted molecular weight: 42 kDa.
ICC/IF
1/50 - 1/100.
IP
1/30.
Notes
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Predicted molecular weight: 42 kDa.
ICC/IF
1/50 - 1/100.
IP
1/30.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function

      Constitutes one of the E3 ubiquitin-protein ligases that mediate 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. 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. Compared to RNF2/RING2, it does not have the main E3 ubiquitin ligase activity on histone H2A, and it may rather act as a modulator of RNF2/RING2 activity.
    • Pathway

      Protein modification; protein ubiquitination.
    • Sequence similarities

      Contains 1 RING-type zinc finger.
    • Cellular localization

      Nucleus speckle.
    • Target information above from: UniProt accession Q06587 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 462604 Chimpanzee
      • Entrez Gene: 101143477 Gorilla
      • Entrez Gene: 6015 Human
      • Omim: 602045 Human
      • SwissProt: A2T6X5 Chimpanzee
      • SwissProt: Q5TJF3 Dog
      • SwissProt: A1YER5 Gorilla
      • SwissProt: Q06587 Human
      • SwissProt: Q8WMN5 Rhesus monkey
      • Unigene: 727565 Human
      see all
    • Alternative names

      • Ring1A antibody
      • E3 ubiquitin-protein ligase RING1 antibody
      • Polycomb complex protein RING1 antibody
      • Really interesting new gene 1 protein antibody
      • RING finger protein 1 antibody
      • Ring1 antibody
      • RING1_HUMAN antibody
      • RING1A antibody
      • Rnf1 antibody
      • Transcription repressor Ring1A antibody
      see all

    Images

    • Immunoprecipitation - Anti-RING1 antibody [EPR13047] (ab180170)
      Immunoprecipitation - Anti-RING1 antibody [EPR13047] (ab180170)
      RING1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab180170 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab180170 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/10000 dilution.

      Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

      Lane 2: ab180170 IP in HeLa whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab180170 in mouse cerebellum tissue lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 15 seconds

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] (ab180170)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] (ab180170)

      Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling RING1 using ab180170 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-RING1 antibody [EPR13047] (ab180170)
      Western blot - Anti-RING1 antibody [EPR13047] (ab180170)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: RING1 knockout HAP1 cell lysate (20 µg)
      Lane 3: MOLT4 cell lysate (20 µg)
      Lane 4: LnCaP cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab180170 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab180170 was shown to specifically react with RING1 when RING1 knockout samples were used. Wild-type and RING1 knockout samples were subjected to SDS-PAGE. ab180170 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-RING1 antibody [EPR13047] (ab180170)
      Western blot - Anti-RING1 antibody [EPR13047] (ab180170)
      All lanes : Anti-RING1 antibody [EPR13047] (ab180170) at 1/5000 dilution

      Lane 1 : Ramos (human Burkitt's lymphoma) whole cell lysate
      Lane 2 : Molt-4 (human acute lymphoblastic leukemia) whole cell lysate
      Lane 3 : LnCaP (human prostat carcinoma) whole cell lysate
      Lane 4 : HeLa (human cervix adenocarcinoma) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/2000 dilution

      Predicted band size: 42 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration 5% NFDM/TBST.

      Exposure Time:
      Lane 1-3: 3 minutes;
      Lane 4: 1 minute

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] (ab180170)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] (ab180170)

      Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling RING1 using ab180170 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-RING1 antibody [EPR13047] (ab180170)
      Immunocytochemistry/ Immunofluorescence - Anti-RING1 antibody [EPR13047] (ab180170)

      Immunofluorescence analysis of HeLa cells labeling RING1 using ab180170 at 1/50 dilution.

    • Anti-RING1 antibody [EPR13047] (ab180170)
      Anti-RING1 antibody [EPR13047] (ab180170)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

    Publishing research using ab180170? Please let us know so that we can cite the reference in this datasheet.

    ab180170 has not yet been referenced specifically in any publications.

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