Recombinant
RabMAb

Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524)

Overview

  • Product name
    Anti-ARF1 + ARF3 antibody [EP442Y]
  • Description
    Rabbit monoclonal [EP442Y] to ARF1 + ARF3
  • Specificity
    The immunogen used for this product shares 100% homology with ARF3, ARF4 and ARF5. Cross-reactivity with the latter two protein have not been confirmed experimentally.
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues in human ARF1.

  • Positive control
    • HEK-293 cell lysates and human breast carcinoma tissue.
  • 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

Applications

Our Abpromise guarantee covers the use of ab32524 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 1/2000. Detects a band of approximately 20 kDa (predicted molecular weight: 21 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for ICC or IP.
  • Target

    Images

    • All lanes : Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524) at 1/20000 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
      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 )

       

    • Immunofluorescence staining of MCF7 cells with purified ab32524 working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 100% methanol. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

    • Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524) at 1/2000 dilution + HEK-293 cell lysate

      Predicted band size : 21 kDa
      Observed band size : 20 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab32524 at 1/250 dilution.
    • Overlay histogram showing HeLa cells stained with ab32524 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32524, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    References

    This product has been referenced in:
    • Covington JD  et al. Skeletal muscle perilipin 3 and coatomer proteins are increased following exercise and are associated with fat oxidation. PLoS One 9:e91675 (2014). WB ; Human . Read more (PubMed: 24632837) »
    • 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 . Read more (PubMed: 24792215) »

    See all 6 Publications for this product

    Customer reviews and Q&As

    Thank you for your enquiry.

    I contacted the laboratories regarding the unsuitability of this antibody for ICC-IF. When it was tested, it showed incorrect staining localization. It worked well in the other applications listed on the datasheet....

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    Thank you for your enquiry. The peptide sequence for this product is:CNTQGLIFVVDSNDRER I am sorry but we do not have any information on whether this product will also detect other ARFs but I can confirm that this antibody recognizes ARF1 (small ...

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    Thank you for your enquiry. This antibody should work very well on your HEK293 cells, since this cell type has been shown to express this protein at detectable amounts. Since this cell type is listed as the positive control, you should be able to s...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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