RecombinantRabMAb

Recombinant Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)

Submit a review Submit a question References (4)
Western blot - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
  • Immunocytochemistry/ Immunofluorescence - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
  • Western blot - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
  • Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR443] to ARF1 + ARF3
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ARF1 + ARF3 antibody [EPR443]
    See all ARF1+ARF3 primary antibodies

  • Description

    Rabbit monoclonal [EPR443] to ARF1 + ARF3

  • Host species

    Rabbit

  • Specificity

    The immunogen used to raise this antibody shares 100% homology with human ARF3, 96% homology with ARF4 and ARF5, and 88% homology with ARF6. Cross-reactivity with the latter three proteins have not been confirmed experimentally.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human ARF1 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • Human fetal kidney, Human fetal brain and MCF7 lysates; Human breast ductal carcinoma, and Human colonic adenocarcinoma tissues This antibody gave a positive result when used in the following methanol fixed cell lines: CaCo2

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab108347 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 - 1/10000. Predicted molecular weight: 21 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/50 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt or IP.

    • Target

    Images

    • Western blot - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      Western blot - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      All lanes : Anti-ARF1 + ARF3 antibody [EPR443] (ab108347) at 1/50000 dilution

      Lane 1 : Full length human ARF-1 recombinant protein
      Lane 2 : Full length human ARF-3 recombinant protein

      Lysates/proteins at 0.015 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 21 kDa
      Observed band size: 23 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and diluting buffer: 5% NFDM /TBST

      Human ARF-1 recombinant protein, (Cat#:ab123538 )
      Human ARF-3 recombinant protein, (Cat#:ab113581 )

    • Immunocytochemistry/ Immunofluorescence - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      Immunocytochemistry/ Immunofluorescence - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)

      ICC/IF image of ab108347 stained CaCo2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab108347 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Western blot - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      Western blot - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      All lanes : Anti-ARF1 + ARF3 antibody [EPR443] (ab108347) at 1/1000 dilution

      Lane 1 : Human fetal kidney lysate
      Lane 2 : Human fetal brain lysate
      Lane 3 : MCF7 lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 21 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)

      Immunohistochemical analysis of ARF1 in paraffin embedded Human breast ductal carcinoma tissue, using ab108347 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)

      Immunohistochemical analysis of ARF1 in paraffin embedded Human colonic adenocarcinoma tissue, using ab108347 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)
      Anti-ARF1 + ARF3 antibody [EPR443] (ab108347)

    References (4)

    Publishing research using ab108347? Please let us know so that we can cite the reference in this datasheet.

    ab108347 has been referenced in 4 publications.

    • Rafiq NB  et al. Podosome assembly is controlled by the GTPase ARF1 and its nucleotide exchange factor ARNO. J Cell Biol 216:181-197 (2017). WB . PubMed: 28007915
    • Eiseler T  et al. Protein Kinase D2 Assembles a Multiprotein Complex at the Trans-Golgi Network to Regulate Matrix Metalloproteinase Secretion. J Biol Chem 291:462-77 (2016). PubMed: 26507660
    • Schultz ML  et al. CLN3 deficient cells display defects in the ARF1-Cdc42 pathway and actin-dependent events. PLoS One 9:e96647 (2014). WB ; Mouse . PubMed: 24792215
    • Jung JJ  et al. Secretion of soluble vascular endothelial growth factor receptor 1 (sVEGFR1/sFlt1) requires Arf1, Arf6, and Rab11 GTPases. PLoS One 7:e44572 (2012). PubMed: 22962618

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