Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

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Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Flow Cytometry - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
  • Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14873] to RAB8A - Low endotoxin, Azide free
  • Suitable for: Flow Cyt, ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free
    See all RAB8A primary antibodies

  • Description

    Rabbit monoclonal [EPR14873] to RAB8A - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human RAB8A aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: P61006

  • Positive control

    • WB: HepG2, HeLa, 293T, HCT-116, C6, Raw264.7, PC-12 and NIH3T3 lysates. IHC-P: Human stomach and Human adenocarcinoma of colon tissues. ICC/IF: HepG2 cells. Flow Cyt: HeLa cells.

  • General notes

    ab221780 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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 ab221780 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 Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    May be involved in vesicular trafficking and neurotransmitter release. Together with RAB11A, RAB3IP, the exocyst complex, PARD3, PRKCI, ANXA2, CDC42 and DNMBP promotes transcytosis of PODXL to the apical membrane initiation sites (AMIS), apical surface formation and lumenogenesis. Together with MYO5B and RAB11A participates in epithelial cell polarization.

  • Sequence similarities

    Belongs to the small GTPase superfamily. Rab family.

  • Cellular localization

    Cell membrane. Golgi apparatus. Cytoplasm > perinuclear region. Cell projection. Colocalizes with OPTN at the Golgi complex and in vesicular structures close to the plasma membrane. In the GDP-bound form, present in the perinuclear region. Shows a polarized distribution to distal regions of cell protrusions in the GTP-bound form. Colocalizes with PARD3, PRKCI, EXOC5, OCLN, PODXL and RAB11A in apical membrane initiation sites (AMIS) during the generation of apical surface and luminogenesis.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AA409338 antibody
    • MEL antibody
    • Mel transforming oncogene (derived from cell line NK14) RAB8 homolog antibody
    • Mel transforming oncogene (derived from cell line NK14) antibody
    • Mel transforming oncogene (RAB8 homolog) antibody
    • Mel transforming oncogene antibody
    • MGC124948 antibody
    • Oncogene c mel antibody
    • Oncogene c-mel antibody
    • RAB8 antibody
    • RAB8A antibody
    • RAB8A member RAS oncogene family antibody
    • RAB8A_HUMAN antibody
    • Ras associated protein RAB8 antibody
    • Ras related protein Rab 8A antibody
    • Ras-related protein Rab-8A antibody
    see all

Images

  • Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    All lanes : Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : RAB8A knockout HeLa cell lysate
    Lane 3 : HCT116 cell lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab188574).

    Lanes 1-4: Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab188574 Anti-RAB8A antibody [EPR14873] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 10000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    All lanes : Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : RAB8A knockout HeLa cell lysate
    Lane 3 : HCT116 cell lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab188574).

    Lanes 1-4: Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab188574 Anti-RAB8A antibody [EPR14873] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264994 (knockout cell lysate ab257196) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 10000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

    This WB data was generated using the same anti-RAB8A antibody clone, EPR14873, in a different buffer formulation (cat# ab188574).

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: RAB8A knockout HAP1 cell lysate (20 µg) 
    Lane 3: HCT116 cell lysate (20 µg)
    Lane 4: NIH/3T3 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab188574 was shown to specifically react with RAB8A when RAB8A knockout samples were used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • Flow Cytometry - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Flow Cytometry - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

    Flow Cytometrical analysis of HeLa cells labeling RAB8A with ab188574 at 1/260 (pink) compared to a Isotype control (green). FITC-conjugated goat-anti-rabbit secondary antibody at 1/150 were used for the analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188574).

  • Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed HepG2 cells labelling RAB8A with ab188574 at 1/500 followed by Goat anti rabbit IgG (Alexa Fluor® 488) and DAPI staining.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188574).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

    Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling RAB8A with ab188574 at 1/500. A prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188574).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

    This IHC data was generated using the same anti-RAB8A antibody clone, EPR14873, in a different buffer formulation (cat# ab188574).

    Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of colon tissue labelling RAB8A with ab188574 at 1/500. A prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)
    Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (ab221780)

References (2)

Publishing research using ab221780? Please let us know so that we can cite the reference in this datasheet.

ab221780 has been referenced in 2 publications.

  • Burikhanov R  et al. Chloroquine-Inducible Par-4 Secretion Is Essential for Tumor Cell Apoptosis and Inhibition of Metastasis. Cell Rep 18:508-519 (2017). PubMed: 28076793
  • Vanhoutte D  et al. Thrombospondin expression in myofibers stabilizes muscle membranes. Elife 5:N/A (2016). WB ; Mouse . PubMed: 27669143

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