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Validated using a knockout cell lineRecombinantRabMAb

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

Submit a review Submit a question References (1)
Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Spa-1 antibody [EPR14134] (ab189929)
  • Immunocytochemistry/ Immunofluorescence - Anti-Spa-1 antibody [EPR14134] (ab189929)
  • Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
  • Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)
  • Flow Cytometry (Intracellular) - Anti-Spa-1 antibody [EPR14134] (ab189929)
  • Anti-Spa-1 antibody [EPR14134] (ab189929)

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: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 (intra): 293T cells.

  • 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

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/400.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
Notes
Flow Cyt (Intra)
1/400.

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

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

      Intracellular flow cytometric analysis of 293T cells fixed in 2% paraformaldehyde labeling Spa-1 with ab189929 at 1/400 dilution,andGoat 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 (1)

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

    ab189929 has been referenced in 1 publication.

    • Lu A  et al. 5-Aza-2'-deoxycytidine advances the epithelial-mesenchymal transition of breast cancer cells by demethylating Sipa1 promoter-proximal elements. J Cell Sci 133:N/A (2020). PubMed: 32193333

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