Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-RALA antibody [EPR6468] (ab126627)

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Western blot - Anti-RALA antibody [EPR6468] (ab126627)
  • Western blot - Anti-RALA antibody [EPR6468] (ab126627)
  • Flow Cytometry - Anti-RALA antibody [EPR6468] (ab126627)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RALA antibody [EPR6468] (ab126627)
  • Anti-RALA antibody [EPR6468] (ab126627)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6468] to RALA
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-RALA antibody [EPR6468]
    See all RALA primary antibodies

  • Description

    Rabbit monoclonal [EPR6468] to RALA

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human RALA aa 100-200. The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, MCF7, MOLT4, BxPC3 and HepG2 whole cell lysate (ab7900). Flow Cyt: MCF7 cells. IHC-P: Human breast carcinoma tissue.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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 ab126627 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: 24 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
1/1000 - 1/10000.

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

 
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 24 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
1/1000 - 1/10000.

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

 
  • Application notes
    Is unsuitable for ICC/IF or IP.

    • Target

    • Function

      Multifuntional GTPase involved in a variety of cellular processes including gene expression, cell migration, cell proliferation, oncogenic transformation and membrane trafficking. Accomplishes its multiple functions by interacting with distinct downstream effectors. Acts as a GTP sensor for GTP-dependent exocytosis of dense core vesicles. Plays a role in the early stages of cytokinesis and is required to tether the exocyst to the cytokinetic furrow. The RALA-exocyst complex regulates integrin-dependent membrane raft exocytosis and growth signaling. Key regulator of LPAR1 signaling and competes with ADRBK1 for binding to LPAR1 thus affecting the signaling properties of the receptor. Required for anchorage-independent proliferation of transformed cells.

    • Sequence similarities

      Belongs to the small GTPase superfamily. Ras family.

    • Post-translational
      modifications

      Prenylation is essential for membrane localization. The geranylgeranylated form and the farnesylated mutant does not undergo alternative prenylation in response to geranylgeranyltransferase I inhibitors (GGTIs) and farnesyltransferase I inhibitors (FTIs).

    • Cellular localization

      Cell surface. Cell membrane. Cleavage furrow. Midbody. Prior to LPA treatment found predominantly at the cell surface and in the presence of LPA co-localizes with LPAR1 and LPAR2 in the endocytic vesicles. During early cytokinesis localizes at the cleavage furrow membrane. Colocalizes with EXOC2 at the early midbody ring and persists there till maturation of the midbody.
    • Information by UniProt

    • Database links

      • Alternative names

        • MGC48949 antibody
        • Ral a antibody
        • Ral A protein antibody
        • RAL antibody
        • RALA antibody
        • RALA_HUMAN antibody
        • Ras family small GTP binding protein RALA antibody
        • RAS like protein A antibody
        • Ras related protein RalA antibody
        • Ras-related protein Ral-A antibody
        • v ral simian leukemia viral oncogene homolog A (ras related) antibody
        • v ral simian leukemia viral oncogene homolog A antibody
        see all

      Images

      • Western blot - Anti-RALA antibody [EPR6468] (ab126627)
        Western blot - Anti-RALA antibody [EPR6468] (ab126627)
        All lanes : Anti-RALA antibody [EPR6468] (ab126627) at 1/1000 dilution

        Lane 1 : Wild-type HeLa cell lysate
        Lane 2 : RALA knockout HeLa cell lysate
        Lane 3 : MCF7 cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 24 kDa



        Lanes 1-3: Merged signal (red and green). Green - ab126627 observed at 25 kDa. Red - loading control ab7291 observed at 50 kDa.

         ab126627 Anti-RALA antibody [EPR6468] was shown to specifically react with RALA in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265092 (knockout cell lysate ab258165) was used. Wild-type and RALA knockout samples were subjected to SDS-PAGE. ab126627 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 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.

         

      • Western blot - Anti-RALA antibody [EPR6468] (ab126627)
        Western blot - Anti-RALA antibody [EPR6468] (ab126627)
        All lanes : Anti-RALA antibody [EPR6468] (ab126627) at 1/1000 dilution

        Lane 1 : MCF7 cell lysate
        Lane 2 : MOLT4 cell lysate
        Lane 3 : BxPC3 cell lysate
        Lane 4 : HepG2 cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

        Predicted band size: 24 kDa

      • Flow Cytometry - Anti-RALA antibody [EPR6468] (ab126627)
        Flow Cytometry - Anti-RALA antibody [EPR6468] (ab126627)
        Overlay histogram showing MCF7 cells stained with ab126627 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab126627, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RALA antibody [EPR6468] (ab126627)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RALA antibody [EPR6468] (ab126627)

        ab126627, at 1/100, staining RALA in formalin fixed, paraffin embedded Human breast carcinoma tissue by Immunohistochemistry.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Anti-RALA antibody [EPR6468] (ab126627)
        Anti-RALA antibody [EPR6468] (ab126627)

      References (4)

      Publishing research using ab126627? Please let us know so that we can cite the reference in this datasheet.

      ab126627 has been referenced in 4 publications.

      • Lee SH & Mayr C Gain of Additional BIRC3 Protein Functions through 3'-UTR-Mediated Protein Complex Formation. Mol Cell 74:701-712.e9 (2019). PubMed: 30948266
      • Chen Q  et al. Subcellular localization of aquaporin 3 in prostate cancer is regulated by RalA. Oncol Rep 39:2171-2177 (2018). PubMed: 29532894
      • Omsland M  et al. Tunneling nanotube (TNT) formation is downregulated by cytarabine and NF-?B inhibition in acute myeloid leukemia (AML). Oncotarget 8:7946-7963 (2017). PubMed: 27974700
      • Lianoglou S  et al. Ubiquitously transcribed genes use alternative polyadenylation to achieve tissue-specific expression. Genes Dev 27:2380-96 (2013). WB ; Human . PubMed: 24145798

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