Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Spa-1 antibody [EPR14134] (ab189929)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14134] to Spa-1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Spa-1 antibody [EPR14134]
    See all Spa-1 primary antibodies

  • Description

    Rabbit monoclonal [EPR14134] to Spa-1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    within Human Spa-1 aa 950 to the C-terminus. The exact sequence is proprietary.
    Database link: Q96FS4

  • Positive control

    • WB: Human fetal thymus, HeLa, Raji, HEK-293T and Raw 264.7 lysates. IHC-P: Human kidney tissue. ICC/IF: A431 cells. Flow Cyt: 293T cells.

  • General notes

     

     

    Previously labelled as SIPA1. 

    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.

    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 ab189929 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 - 1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 112 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.
ICC/IF 1/250 - 1/500.
Flow Cyt 1/400.

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

 

Target

  • Function

    GTPase activator for the nuclear Ras-related regulatory proteins Rap1 and Rap2 in vitro, converting it to the putatively inactive GDP-bound state.

  • Tissue specificity

    Expressed in fetal as well as in adult tissues. Expressed abundantly in the lymphoid tissues such as thymus, spleen and peripheral blood lymphocytes and also shows a significant expression in the spinal cord.

  • Sequence similarities

    Contains 1 PDZ (DHR) domain.
    Contains 1 Rap-GAP domain.

  • Cellular localization

    Nucleus. Cytoplasm > perinuclear region. Endomembrane system. Mostly localized in the perinuclear membraneous region.
  • Information by UniProt

  • Database links

    • Alternative names

      • GTPase activating protein Spa 1 antibody
      • GTPase-activating protein Spa-1 antibody
      • MGC102688 antibody
      • MGC17037 antibody
      • p130 SPA-1 antibody
      • p130 SPA1 antibody
      • Signal induced proliferation associated 1 antibody
      • Signal induced proliferation associated gene 1 antibody
      • Signal induced proliferation associated protein 1 antibody
      • Signal-induced proliferation-associated protein 1 antibody
      • SIPA 1 antibody
      • Sipa-1 antibody
      • Sipa1 antibody
      • SIPA1_HUMAN antibody
      • Spa1 antibody
      see all

    Images

    • Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
      Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
      All lanes : Anti-Spa-1 antibody [EPR14134] (ab189929) at 1/5000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : Spa-1 knockout HeLa cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : HEK-293 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 112 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab189929 observed at 130 kDa. Red - loading control ab7291 observed at 50 kDa.

       ab189929 Anti-Spa-1 antibody [EPR14134] was shown to specifically react with Spa-1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265434 (knockout cell lysate ab258189) was used. Wild-type and Spa-1 knockout samples were subjected to SDS-PAGE. ab189929 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 5000 dilution and 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-Spa-1 antibody [EPR14134] (ab189929)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Spa-1 antibody [EPR14134] (ab189929)

      Immunohistochemical analysis of formalin fixed, paraffin embedded Human kidney tissue labeling Spa-1 with ab189929 at 1/50 dilution, and Prediluted HRP Polymer for Rabbit IgG as a secondary antibody. Counterstained with Hematoxylin.

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

    • Immunocytochemistry/ Immunofluorescence - Anti-Spa-1 antibody [EPR14134] (ab189929)
      Immunocytochemistry/ Immunofluorescence - Anti-Spa-1 antibody [EPR14134] (ab189929)

      Immunocytochemical analysis of A431 cells fixed in 4% paraformaldehyde labeling Spa-1 with ab189929 at 1/250 dilution, and Goat anti rabbit IgG(Alexa Fluor® 555) at 1/200 dilution as the secondary antibody. Counterstained with DAPI.

    • Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
      Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
      All lanes : Anti-Spa-1 antibody [EPR14134] (ab189929) at 1/5000 dilution

      Lane 1 : Human fetal thymus
      Lane 2 : Raji cell lysate
      Lane 3 : 293T 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: 112 kDa
      Additional bands at: 130 kDa. We are unsure as to the identity of these extra bands.

    • Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
      Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
      Anti-Spa-1 antibody [EPR14134] (ab189929) at 1/1000 dilution + Raw264.7 at 10 µg

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

      Predicted band size: 112 kDa
      Additional bands at: 130 kDa. We are unsure as to the identity of these extra bands.

    • Flow Cytometry - Anti-Spa-1 antibody [EPR14134] (ab189929)
      Flow Cytometry - Anti-Spa-1 antibody [EPR14134] (ab189929)

      Flow cytometric analysis of 293T cells fixed in 2% paraformaldehyde labeling Spa-1 with ab189929 at 1/400 dilution, and Goat anti rabbit IgG (FITC) at 1/150 dilution as the secondary antibody.

    • Anti-Spa-1 antibody [EPR14134] (ab189929)
      Anti-Spa-1 antibody [EPR14134] (ab189929)

    References (0)

    Publishing research using ab189929? Please let us know so that we can cite the reference in this datasheet.

    ab189929 has not yet been referenced specifically in any publications.

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