Overview

  • Product name
    Anti-RhoA antibody [EPR18134]
    See all RhoA primary antibodies
  • Description
    Rabbit monoclonal [EPR18134] to RhoA
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human RhoA aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: P61586

  • Positive control
    • WB: Human RhoA full length protein; HeLa, HEK-293 C6, Raw264.7 and NIH/3T3 cell lysates, human fetal brain and fetal kidney lysates, mouse brain, kidney and spleen lysates, rat brain, kidney and spleen lysates. ICC/IF: Jurkat and K562 cells. Flow Cyt: HeLa cells.
  • General notes

    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 ab187027 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 1/5000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 28671996
ICC/IF 1/150.
Flow Cyt 1/200.

Target

  • Function
    Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague, and Yersinia pseudotuberculosis, which causes gastrointestinal disorders. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP.
  • Sequence similarities
    Belongs to the small GTPase superfamily. Rho family.
  • Domain
    The basic-rich region is essential for yopT recognition and cleavage.
  • Post-translational
    modifications
    Substrate for botulinum ADP-ribosyltransferase.
    Cleaved by yopT protease when the cell is infected by some Yersinia pathogens. This removes the lipid attachment, and leads to its displacement from plasma membrane and to subsequent cytoskeleton cleavage.
    AMPylation at Tyr-34 and Thr-37 are mediated by bacterial enzymes in case of infection by H.somnus and V.parahaemolyticus, respectively. AMPylation occurs in the effector region and leads to inactivation of the GTPase activity by preventing the interaction with downstream effectors, thereby inhibiting actin assembly in infected cells. It is unclear whether some human enzyme mediates AMPylation; FICD has such ability in vitro but additional experiments remain to be done to confirm results in vivo.
    Ubiquitinated by the BCR(BACURD1) and BCR(BACURD2) E3 ubiquitin ligase complexes, leading to its degradation by the proteasome, thereby regulating the actin cytoskeleton and cell migration.
  • Cellular localization
    Cell membrane. Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Aplysia ras related homolog 12 antibody
    • ARH12 antibody
    • ARHA antibody
    • H 12 antibody
    • H12 antibody
    • Oncogene RHO H12 antibody
    • Ras homolog family member A antibody
    • Ras homolog gene family member A antibody
    • Rho A antibody
    • Rho cDNA clone 12 antibody
    • RHO H12 antibody
    • RHO12 antibody
    • RHOA antibody
    • RHOA_HUMAN antibody
    • RHOH12 antibody
    • Small GTP binding protein Rho A antibody
    • Transforming protein Rho A antibody
    • Transforming protein RhoA antibody
    see all

Images

  • All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/1000 dilution

    Lane 1 : Wild-type Hek293T whole cell lysate
    Lane 2 : RHOA knockout Hek293T whole cell lysate
    Lane 3 : Jurkat whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 22 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab187027 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab187027 was shown to specifically react with RhoA in wild-type Hek293T cells as signal was lost in RHOA knockout cells. Wild-type and RHOA knockout samples were subjected to SDS-PAGE. Ab187027 and ab9484 (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% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling RhoA with ab187027 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Jurkat cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab187027 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

    Lane 1 : Human RhoA full length protein
    Lane 2 : Human RhoB full length protein
    Lane 3 : Human RhoC full length protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

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


    Exposure time: 1 second


    5% NFDM/TBST: Blocking and diluting buffer.

    Human RhoA full length protein (ab101594) contains aa1-193 with His-Tag® at N-Terminus; Human RhoB full length protein (ab107139) contains aa1-193 with His-Tag® at N-Terminus; Human RhoC full length protein (ab98085) contains aa1-190 with His-Tag® at N-Terminus.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling RhoA with ab187027 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and a unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Photoreceptor expression of RhoA and Cdc42.

    Retinal central cross sections stained with RhoA (A, red) and counterstained with DAPI (blue). Both strains showed minimal RhoA staining in the photoreceptors and inner nuclear layer in room air controls (RA) and HIPR. There were no distinct differences detected at any time point at the level of the photoreceptors. At P21 in HIPR in both strains, RhoA was increased in the Mϋller cells, compared to respective room controls. Representative images from the central retina are shown (N = 3). Scale bar, 50 μm. ONL, outer nuclear layer. RPE, retinal pigment epithelium.

  • All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

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


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat kidney lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 8 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

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


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling RhoA with ab187027 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab187027 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

References

This product has been referenced in:
  • Zhao F  et al. Exendin-4 promotes actin cytoskeleton rearrangement and protects cells from Nogo-A-?20 mediated spreading inhibition and growth cone collapse by down-regulating RhoA expression and activation via the PI3K pathway. Biomed Pharmacother 109:135-143 (2019). Read more (PubMed: 30396070) »
  • Kataoka Y  et al. Hypoxia-induced galectin-3 enhances RhoA function to activate the motility of tumor cells in non-small cell lung cancer. Oncol Rep 41:853-862 (2019). Read more (PubMed: 30535445) »
See all 20 Publications for this product

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