Overview

  • Product name
    Anti-RhoA antibody [1B12]
    See all RhoA primary antibodies
  • Description
    Mouse monoclonal [1B12] to RhoA
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein, corresponding to amino acids 1-194 of Human RhoA

  • Positive control
    • HeLa cells HL-60 cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab54835 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 a concentration of 1 - 5 µg/ml. Predicted molecular weight: 22 kDa.
IHC-P Use a concentration of 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 10 µg/ml.
ELISA Use at an assay dependent concentration.

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

Anti-RhoA antibody [1B12] images



  • Predicted band size : 22 kDa

    RhoA antibody (ab54835) at 1ug/lane + HL-60 cell lysate (ab7914) at 25ug/lane.

  • All lanes : Anti-RhoA antibody [1B12] (ab54835) at 1 µg

    Lane 1 : RHOA transfected 293T lysate
    Lane 2 : Non-transfected lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat anti-mouse IgG

    Predicted band size : 22 kDa
  • ICC/IF image of ab54835 stained HeLa cells (ab150035). The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab54835, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells (ab150035) stained with ab54835 (red line). The cells were fixed with methanol (5 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 (ab54835, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • RhoA antibody (ab54835) used in immunohistochemistry at 5ug/ml on formalin fixed and paraffin embedded human lymphoma tissue.
  • ab54835 at 10 ug/ml staining RhoA in human Hela cells (ab150035) by Immunocytochemistry / Immunofluorescence.

References for Anti-RhoA antibody [1B12] (ab54835)

This product has been referenced in:
  • Chang HR  et al. Systematic approach identifies RHOA as a potential biomarker therapeutic target for Asian gastric cancer. Oncotarget 7:81435-81451 (2016). WB ; Human . Read more (PubMed: 27806312) »
  • Han Q  et al. Human beta-defensin-1 suppresses tumor migration and invasion and is an independent predictor for survival of oral squamous cell carcinoma patients. PLoS One 9:e91867 (2014). WB ; Human . Read more (PubMed: 24658581) »

See all 11 Publications for this product

Product Wall

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Hela, HT-60)
Specification
Hela, HT-60
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Feb 11 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step
Enzymatic
Sample
Rat Tissue sections (Gonad)
Specification
Gonad
Permeabilization
No
Fixative
Formaldehyde
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Submitted Sep 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (glioblastoma cells)
Specification
glioblastoma cells
Fixative
Formaldehyde
Permeabilization
Yes - 0,1% TrintonX100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
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Submitted Dec 04 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma cells)
Loading amount
50 µg
Specification
Glioblastoma cells
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Dec 04 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293)
Loading amount
25 µg
Specification
HEK 293
Gel Running Conditions
Non-reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Nov 07 2011

Application
Western blot
Sample
Rat Cell lysate - whole cell (C6 Rat glioma cell line and Rat primary Glia cells)
Loading amount
40 µg
Specification
C6 Rat glioma cell line and Rat primary Glia cells
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Ms. Revital Knirsh

Verified customer

Submitted Jun 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (hela)
Loading amount
20000 cells
Specification
hela
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Jul 01 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HeLa)
Loading amount
25 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (4-15% tris-hcl)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Apr 01 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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