Overview

  • Product name

    Anti-RAB10 antibody [MJF-R23]
    See all RAB10 primary antibodies
  • Description

    Rabbit monoclonal [MJF-R23] to RAB10
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, Flow Cyt, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein corresponding to Human RAB10 aa 1 to the C-terminus. Antibody epitope is C-terminal amino acids DISSGGGVT (AA 185-193)
    Database link: P61026

  • Positive control

    • WB: A549, HeLa, HCT 116, MCF7, NIH/3T3, PC-12 and C6 whole cell lysates. ICC/IF: A549 and MCF7 cells. Flow: A549 cells. IP: A549 whole cell lysate.
  • General notes

    This antibody was developed with support from The Michael J. Fox Foundation.

     

    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 ab237703 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
IP 1/30.
Flow Cyt 1/600.
ICC/IF 1/500.
WB 1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).

Target

  • Function

    The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different set of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion (By similarity). That Rab is mainly involved in the biosynthetic transport of proteins from the Golgi to the plasma membrane. Regulates, for instance, SLC2A4/GLUT4 glucose transporter-enriched vesicles delivery to the plasma membrane. In parallel, it regulates the transport of TLR4, a toll-like receptor to the plasma membrane and therefore may be important for innate immune response. Plays also a specific role in asymmetric protein transport to the plasma membrane within the polarized neuron and epithelial cells. In neurons, it is involved in axonogenesis through regulation of vesicular membrane trafficking toward the axonal plasma membrane while in epithelial cells, it regulates transport from the Golgi to the basolateral membrane. Moreover, may play a role in the basolateral recycling pathway and in phagosome maturation. According to PubMed:23263280, may play a role in endoplasmic reticulum dynamics and morphology controlling tubulation along microtubules and tubules fusion.
  • Sequence similarities

    Belongs to the small GTPase superfamily. Rab family.
  • Cellular localization

    Cytoplasmic vesicle membrane. Golgi apparatus membrane. Golgi apparatus, trans-Golgi network membrane. Endosome membrane. Recycling endosome membrane. Cytoplasmic vesicle, phagosome membrane. Cell projection, cilium. Endoplasmic reticulum membrane. Associates with SLC2A4/GLUT4 storage vesicles (PubMed:22908308). Localizes to the base of the cilium (PubMed:20576682). Transiently associates with phagosomes (By similarity). Localizes to the endoplasmic reticulum at domains of new tubule growth (PubMed:23263280).
  • Information by UniProt
  • Database links

  • Alternative names

    • GTP binding protein RAB10 antibody
    • Rab10 antibody
    • RAB10 member RAS oncogene family antibody
    • RAB10_HUMAN antibody
    • Ras related GTP binding protein antibody
    • Ras related GTP binding protein RAB10 antibody
    • Ras-related protein Rab-10 antibody
    see all

Images

  • All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution

    Lane 1 : Wild-type A549 whole cell lysate
    Lane 2 : RAB10 knockout A549 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 22 kDa
    Observed band size: 25 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab237703 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab237703 was shown to recognize RAB10 in wild-type A549 cells as signal was lost at the expected MW in RAB10 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RAB10 knockout samples were subjected to SDS-PAGE. Ab237703 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue). 

  • All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution

    Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell line) whole cell lysate
    Lane 2 : Rab10 knockout A549 whole cell lysate
    Lane 3 : HCT 116 (human colorectal carcinoma epithelial cell line) whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
    Lane 6 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 7 : C6 (rat glial tumor glial cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 22 kDa
    Observed band size: 22 kDa


    Exposure time: 10 seconds


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST 

    The WT and Rab10 KO A549 lysates were kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.

  • RAB10 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma cell line) whole cell lysate using ab237703 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab237703 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution was used for detection.

    Lane 1: A549 whole cell lysate 10μg (input)
    Lane 2: ab237703 IP in A549 whole cell lysate.
    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab237703 in A549 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in MCF7 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in A549 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 with ab237703 at 1/600 dilution (red) compared with the rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue). 

References

ab237703 has not yet been referenced specifically in any publications.

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