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  1. Link

    ufc1-antibody-epr15014-102-ab189252.pdf

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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ubiquitin E2 Enzymes
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-UFC1 antibody [EPR15014-102] (ab189252)

  • Datasheet
  • Certificate of Compliance
Reviews (1) Submit a question References (2)

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Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Immunoprecipitation - Anti-UFC1 antibody [EPR15014-102] (ab189252)
  • Anti-UFC1 antibody [EPR15014-102] (ab189252)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15014-102] to UFC1
  • Suitable for: IP, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-UFC1 antibody [EPR15014-102]
    See all UFC1 primary antibodies
  • Description

    Rabbit monoclonal [EPR15014-102] to UFC1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WB, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal liver tissue lysate and HEK-293T, HAP1, U87-MG, MCF7, A549, RAW 264.7, and NIH 3T3 cell lysates. IHC-P: Human colon and human transitional cell carcinoma of bladder tissues.
  • 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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR15014-102
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E2 Enzymes

Associated products

  • Alternative Versions

    • Anti-UFC1 antibody [EPR15014-102] - BSA and Azide free (ab250984)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human UFC1 knockout HEK-293T cell line (ab266814)
  • KO cell lysates

    • Human UFC1 knockout HEK-293T cell lysate (ab257781)
  • Positive Controls

    • MCF7 whole cell lysate (ab29537)
    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
    • A549 whole cell lysate (ab7910)
  • Recombinant Protein

    • Recombinant Human UFC1 protein (ab107151)
  • Related Products

    • Recombinant Human UFC1 protein (ab107151)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab189252 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
IP
1/50.
WB (1)
1/1000 - 1/10000. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
IP
1/50.
WB
1/1000 - 1/10000. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF.

Target

  • Function

    E2-like enzyme which forms an intermediate with UFM1 via a thioester linkage.
  • Sequence similarities

    Belongs to the ubiquitin-conjugating enzyme family. UFC1 subfamily.
  • Target information above from: UniProt accession Q9Y3C8 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 51506 Human
    • Omim: 610554 Human
    • SwissProt: Q9Y3C8 Human
    • Unigene: 301412 Human
    • Alternative names

      • CGI-126 antibody
      • HSPC155 antibody
      • Ubiquitin fold modifier conjugating enzyme 1 antibody
      • Ubiquitin-fold modifier-conjugating enzyme 1 antibody
      • UFC1 antibody
      • UFC1_HUMAN antibody
      • Ufm1 conjugating enzyme 1 antibody
      • Ufm1-conjugating enzyme 1 antibody
      see all

    Images

    • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : UFC1 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 19 kDa
      Observed band size: 20 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab189252 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab189252 was shown to react with UFC1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266814 (knockout cell lysate ab257781) was used. Wild-type HEK-293T and UFC1 HEK-293T KO cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab189252 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon tissue labeling UFC1 with ab189252 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/10000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : UFC1 knockout HAP1 cell lysate
      Lane 3 : MCF7 cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 19 kDa
      Observed band size: 20 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab189252 observed at 20 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab189252 was shown to react with UFC1 in wild-type HAP1 cells in western blot. Loss of signal was observed when UFC1 knockout sample was used. Wild-type HAP1 and UFC1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab189252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/20000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : U87-MG cell lysate
      Lane 3 : MCF7 cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 19 kDa

    • Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/5000 dilution

      Lane 1 : RAW 264.7 cell lysate
      Lane 2 : NIH 3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 19 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling UFC1 with ab189252 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Immunoprecipitation - Anti-UFC1 antibody [EPR15014-102] (ab189252)

      Western blot analysis of immunoprecipitation pellet from MCF7cell lysate immunoprecipitated using ab189252 at 1/50 dilution (lane 1) or PBS control (lane 2).
      Secondary: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

    • Anti-UFC1 antibody [EPR15014-102] (ab189252)
      Anti-UFC1 antibody [EPR15014-102] (ab189252)

    Protocols

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

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

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

    References (2)

    Publishing research using ab189252? Please let us know so that we can cite the reference in this datasheet.

    ab189252 has been referenced in 2 publications.

    • Qin B  et al. STK38 promotes ATM activation by acting as a reader of histone H4 ufmylation. Sci Adv 6:eaax8214 (2020). PubMed: 32537488
    • Qin B  et al. UFL1 promotes histone H4 ufmylation and ATM activation. Nat Commun 10:1242 (2019). PubMed: 30886146

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Western blot abreview for Anti-UFC1 antibody [EPR15014-102]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Gel Running Conditions
    Reduced Denaturing (15)
    Loading amount
    50 µg
    Treatment
    siRNAs for 48-72 h
    Specification
    U2OS cells
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 25 2022

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