Recombinant Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Product name

Anti-RhoA + RhoC antibody [EPR18133]
See all RhoA + RhoC primary antibodies
Description

Rabbit monoclonal [EPR18133] to RhoA + RhoC
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human RhoA full length protein, Human RhoC full length protein. HepG2, HeLa, Jurkat and C6, Raw264.7, PC12 and NIH 3T3 cell lysates. Human fetal brain and fetal kidney lysates; Mouse brain and kidney lysates; Rat brain and kidney lysates. IHC-P: Transitional cell carcinoma of bladder tissue, Mouse kidney and rat stomach tissues. ICC/IF: NIH3T3 cells. Flow Cyt (intra): Jurkat cells. IP: NIH/3T3 cell lysate.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18133
Isotype

IgG
Research areas

Alternative Versions
- PE Anti-RhoA + RhoC antibody [EPR18133] (ab225344)
- Anti-RhoA + RhoC antibody [EPR18133] - BSA and Azide free (ab238943)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab187026 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
Flow Cyt (Intra)		1/400.
WB		1/2000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/500.
IP		1/50.

Notes
Flow Cyt (Intra) 1/400.
WB 1/2000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
IP 1/50.

Relevance

Transforming protein RhoA: Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion. Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague, and Yersinia pseudotuberculosis, which causes gastrointestinal disorders. Stimulates PKN2 kinase activity. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization.Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion. May be an activator of PLCE1. Activated by ARHGEF2, which promotes the exchange of GDP for GTP. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity. In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization (By similarity). Regulates KCNA2 potassium channel activity by reducing its location at the cell surface in response to CHRM1 activation; promotes KCNA2 endocytosis. Rho-related GTP-binding protein RhoC: Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Serves as a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis. Regulates apical junction formation in bronchial epithelial cells.
Database links
- Entrez Gene: 387 Human
- Entrez Gene: 389 Human
- SwissProt: P08134 Human
- SwissProt: P61586 Human
- SwissProt: Q62159 Mouse
- SwissProt: Q9QUI0 Mouse
- SwissProt: P61589 Rat
Alternative names
- ARH12 antibody
- ARH9 antibody
- ARHA antibody
- ARHC antibody
- H9 antibody
- Rho cDNA clone 12 antibody
- Rho cDNA clone 9 antibody
- Rho-related GTP-binding protein RhoC antibody
- RHO12 antibody
- RHOA antibody
- RHOC antibody
- RHOH12 antibody
- RHOH9 antibody
- Transforming protein RhoA antibody
see all

Western blot - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Lanes 1-2 : Anti-RhoA + RhoC antibody [EPR18133] (ab187026) at 1/20000 dilution
Lane 3 : Anti-RhoA + RhoC antibody [EPR18133] (ab187026) at 1/500 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),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 22 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?

5% NFDM/TBST: Blocking and diluting buffer.
Exposure time: Lane 1: 30 seconds; lane 2 and 3: 3 minutes.
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.
The affinity of ab187016 to RhoA is higher than to RhoC.
Western blot - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

All lanes : Anti-RhoA + RhoC antibody [EPR18133] (ab187026) at 1/2000 dilution

Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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

Exposure time: 1 minute

5% NFDM/TBST: Blocking and diluting buffer.
Western blot - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

All lanes : Anti-RhoA + RhoC antibody [EPR18133] (ab187026) at 1/2000 dilution

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

Lysates/proteins at 10 mg/ml 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.
Western blot - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

All lanes : Anti-RhoA + RhoC antibody [EPR18133] (ab187026) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat kidney lysate
Lane 5 : C6 (Rat glial tumor cells) whole cell lysate
Lane 6 : Raw264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus ) whole cell lysate
Lane 7 : PC12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
Lane 8 : NIH 3T3 (Mouse embyro fibroblast cells ) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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

Exposure time: 10 seconds

5% NFDM/TBST: Blocking and diluting buffer.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling RhoA + RhoC with ab187026 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on cancer cells of transitional cell carcinoma of bladder is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling RhoA + RhoC with ab187026 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on kidney tubules of mouse kidney is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling RhoA + RhoC with ab187026 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on epithelial cells of rat stomach is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling RhoA + RhoC with ab187026 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on NIH 3T3 cells 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 1/500 dilution (red).
The negative controls are as follows:
1. ab187026 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Flow Cytometry (Intracellular) - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling RhoA + RhoC with ab187026 at 1/400 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Immunoprecipitation of RhoA + RhoC from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate achieved using ab187026 at 1/50 dilution.
Lane 1: Input: 10µg of NIH/3T3 whole cell lysate.
Lane 2: NIH/3T3 whole cell lysate following IP with ab187026.
Lane 3: Negative control: IP using Rabbit monoclonal IgG (ab172730) instead of ab187026 in NIH/3T3 whole cell lysates.
Western blot was performed using ab187026 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1500 was used for detection.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. 30 second exposure.
Anti-RhoA + RhoC antibody [EPR18133] (ab187026)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Publishing research using ab187026? Please let us know so that we can cite the reference in this datasheet.

ab187026 has been referenced in 1 publication.

Ge Z et al. Overexpression of the hyperplasia suppressor gene inactivates airway fibroblasts obtained from a rat model of chronic obstructive pulmonary disease by inhibiting the Wnt signaling pathway. Mol Med Rep 20:2754-2762 (2019). PubMed: 31322244

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Relevance

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

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