Anti-RhoA (phospho S188) antibody (ab41435)

Overview

  • Product name
    Anti-RhoA (phospho S188) antibody
    See all RhoA primary antibodies
  • Description
    Rabbit polyclonal to RhoA (phospho S188)
  • Specificity
    Phospho-Ser188-RhoA specific. RhoA immunogen sequence not found in RhoB or RhoC proteins.
  • Tested applications
    Suitable for: ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide phosphorylated at Ser188, corresponding to amino acid residues around serine 188 of human RhoA, conjugated to KLH.

  • Positive control
    • Recombinant human RhoA GST fusion protein, phosphorylated with PKA.
  • General notes
    Do not aliquot.


    Post-translational regulation of Rho activity has been shown specifically for RhoA. This Rho protein is phosphorylated in vitro on serine 188 by cAMP- and cGMP-dependent kinases (PKA and PKG). Ser188 phosphorylation enhances RhoGDI binding and inhibits RhoA-mediated stress fiber formation. Thus, Ser188 is an important site for negative regulation of RhoA activity.

Properties

Applications

Our Abpromise guarantee covers the use of ab41435 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
ELISA 1/2000.
WB 1/1000. Predicted molecular weight: 22 kDa.

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

  • Lanes 1 - 2 : anti-Rho
    Lanes 3 - 6 : Anti-RhoA (phospho S188) antibody (ab41435) at 1/1000 dilution

    Lane 1 : recombinant human RhoA GST fusion protein, unphosphorylated
    Lane 2 : recombinant human RhoA GST fusion protein, phosphorylated with PKA
    Lane 3 : recombinant human RhoA GST fusion protein, unphosphorylated
    Lane 4 : recombinant human RhoA GST fusion protein, phosphorylated with PKA
    Lane 5 : recombinant human RhoA GST fusion protein, phosphorylated with PKA with RhoA (phospho S188) peptide
    Lane 6 : recombinant human RhoA GST fusion protein, phosphorylated with PKA with non-specific phosphoserine peptide


    Predicted band size : 22 kDa
    Observed band size : 49 kDa (why is the actual band size different from the predicted?)
    Membrane was incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.

    A fusion protein was used in this western blot; hence the difference in molecular weight compared to predicted molecular weight.

  • All lanes : Anti-RhoA (phospho S188) antibody (ab41435) at 1/1000 dilution

    Lane 1 : MCF7
    Lane 2 : human liver nuclear tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Polyclonal goat anti-rabbit HRP conjugate at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

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


    Exposure time : 25 seconds

    This image is courtesy of an anonymous abreview.

    Blocking: 5% milk for 1 hour at 23°C 

    See Abreview

References

This product has been referenced in:
  • Wang XQ  et al. Resveratrol enhances vascular reactivity in mice following lipopolysaccharide challenge via the RhoA-ROCK-MLCP pathway. Exp Ther Med 14:308-316 (2017). Read more (PubMed: 28672931) »
  • da Silva Almeida AC  et al. The mutational landscape of cutaneous T cell lymphoma and Sézary syndrome. Nat Genet 47:1465-70 (2015). Read more (PubMed: 26551667) »

See all 6 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Rat Cell lysate - whole cell (IMCD cells)
Gel Running Conditions
Reduced Denaturing (12 %)
Loading amount
30 µg
Treatment
- and + 30 ´M forskolin (FSK)
Specification
IMCD cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Mrs. Andrea Geelhaar

Verified customer

Submitted May 19 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - nuclear (liver)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Dec 14 2015

Application
Western blot
Sample
Pig Cell lysate - whole cell (Swine testicle cells)
Loading amount
20 µg
Specification
Swine testicle cells
Gel Running Conditions
Reduced Denaturing (12% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Thary Jacob

Verified customer

Submitted Jan 24 2013

DISCOUNT CODE: ABTRIAL-EBXXX

Expiration date: 17/03/2013
Value: £227.00

I am very pleased to hear you would like to accept our offer and test ab41435 in Pig. This code will give you £227.00 off your next order befo...

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Thank you for your contact.

I am a bit confused about the -mail. Could you explain what you mean by inactive code number?

I look forward to hearing from you soon.

Thank you for your interest.

As it has been described in the previous e-mail the discount code for the value of the product and as soon as the Abreview is submitted, the discount code will become active.

Please do let me know if you...

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Thank you very much for your interest our AbTrial program.


To our knowledge,ab41435 has not been tested in Pig. However by participating in our AbTrial program you can now use our products in an untested application or species without f...

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We have observed both the 27 kDa and the 70 kDa band using RP1361. We have not identified the 70 kDa band, and have no evidence that it is related to RhoA. We have shown that antibody does detect RhoA at the lower molecular weight, as well as detects r...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa Cells)
Loading amount
30 µg
Specification
HeLa Cells
Gel Running Conditions
Non-reduced Denaturing (12.5% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 08 2012

I am very pleased to hear you would like to accept our offer and test ab41435 in mouse in ICC/IF. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for mouse in ICC/IF and include ...

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