Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

Submit a review Submit a question References (2)
Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
  • Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
  • Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
  • Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
  • Flow Cytometry (Intracellular) - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
  • Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18134] to RhoA - Low endotoxin, Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free
    See all RhoA primary antibodies

  • Description

    Rabbit monoclonal [EPR18134] to RhoA - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

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

  • Positive control

    • WB: Human RhoA full length protein; HeLa, HEK293 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 (intra): HeLa cells.

  • General notes

    ab219371 is the carrier-free version of ab187027.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219371 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
Use at an assay dependent concentration. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
ICC/IF
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
ICC/IF
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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

    see all

  • 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

  • Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

    Lanes 1 & 3 : Wild-type HEK-293T cell lysate
    Lanes 2 & 4 : RHOA knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 21 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab187027).

    Lanes 1- 4: Merged signal (red and green). Green - ab187027 observed at 21 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab187027 was shown to react with RhoA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266592 (knockout cell lysate ab257637) was used. Wild-type HEK-293T and RHOA knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187027 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 5000 dilution and a 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    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. The membrane was blocked with 3% Milk. 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Flow Cytometry (Intracellular) - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Flow Cytometry (Intracellular) - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

    Intracellular 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. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027). 

     

     

  • Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

References (2)

Publishing research using ab219371? Please let us know so that we can cite the reference in this datasheet.

ab219371 has been referenced in 2 publications.

  • Tian YM  et al. Elevated expression of the leptin receptor ob-R may contribute to inflammation in patients with ulcerative colitis. Mol Med Rep 20:4706-4712 (2019). PubMed: 31702041
  • Deng M  et al. Silencing cyclin-dependent kinase inhibitor 3 inhibits the migration of breast cancer cell lines. Mol Med Rep 14:1523-30 (2016). WB ; Human . PubMed: 27314680

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

There are currently no Customer reviews or Questions for ab219371.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com