Overview

  • Product nameAnti-RhoA antibody
    See all RhoA primary antibodies
  • Description
    Rabbit polyclonal to RhoA
  • Tested applicationsSuitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Dog, Zebrafish, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human RhoA.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HL60; MCF7; MDA-MB-231; HeLa; Human Platelets.

Properties

Applications

Our Abpromise guarantee covers the use of ab86297 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
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 20 kDa (predicted molecular weight: 21 kDa).

Target

  • FunctionRegulates 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 similaritiesBelongs to the small GTPase superfamily. Rho family.
  • DomainThe 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 localizationCell 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 images

  • All lanes : Anti-RhoA antibody (ab86297) at 1 µg/ml

    Lane 1 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : Human Platelet (Human adult normal cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)


    Exposure time : 90 seconds
  • All lanes : Anti-RhoA antibody (ab86297) at 1 µg/ml

    Lane 1 : Kidney (Mouse) Tissue Lysate
    Lane 2 : Mouse lung normal tissue lysate - total protein (ab29297)
    Lane 3 : Kidney (Rat) Tissue Lysate
    Lane 4 : Lung (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes
  • ICC/IF image of ab86297 stained MCF7 cells. 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 (ab86297, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

References for Anti-RhoA antibody (ab86297)

This product has been referenced in:
  • Freeman MC  et al. Coronaviruses induce entry-independent, continuous macropinocytosis. MBio 5:e01340-14 (2014). Mouse . Read more (PubMed: 25096879) »
  • Nuno DW  et al. RhoA localization with caveolin-1 regulates vascular contractions to serotonin. Am J Physiol Regul Integr Comp Physiol 303:R959-67 (2012). WB ; Mouse . Read more (PubMed: 22955057) »

See all 2 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Uterus)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH=6.0
Permeabilization No
Specification Uterus
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative Formaldehyde
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Verified customer

Submitted Oct 14 2016

Thank you for your reply.

I hope the use of SDS and DTT will resolve the problem!

Please let us know also if there is anything else we could help you with. We do have for example different cell fractionation kits, and if the SDS and...

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Thank you very much for your email. I am very happy to hear that the RhoA antibody is working now.

The observedband in the cytoplasm is indeed unexpected. I would think this would correspond to a complex in the cytoplasm or another modificati...

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Thank you for your call a couple weeks ago, and I sincerely apologize for the delay in getting back to you.
As we discussed, the immunogen of ab86297 is 83% identical to RhoB and 73% to RhoC, so it is possible that this antibody will cross-react. ...

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Thank you for your email.

I look forward hearing back from you with the results of the BSA blocking.

Have a great day!

Thank you for taking time to contact us with this information. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital informati...

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Thank you for contacting Abcam regarding ab86297. We have not tested this antibody for cross reactivity with RhoB or RhoC.  By blasting the immunogen sequence for this Ab, it shows there may be some cross reactivity with RhoC. I hope this infor...

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