Anti-RhoA antibody [1B12] (ab54835)
Key features and details
- Mouse monoclonal [1B12] to RhoA
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-RhoA antibody [1B12]
See all RhoA primary antibodies -
Description
Mouse monoclonal [1B12] to RhoA -
Host species
Mouse -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein corresponding to Human RhoA aa 1-193.
Database link: P61586 -
Positive control
- WB: 293T, HL-60, lysate ICC/IF: HeLa cells. Flow cyt: HeLa cells. IHC-P: Human lymphoma tissue.
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General notes
This product was changed from ascites to tissue culture supernatant on 15 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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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: 7.40
Constituent: 100% PBS -
Concentration information loading...
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Purity
Tissue culture supernatant -
Purification notes
Purified from TCS. -
Clonality
Monoclonal -
Clone number
1B12 -
Isotype
IgG1 -
Light chain type
lambda -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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 |
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WB | (11) |
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa.
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IHC-P | (1) |
Use at an assay dependent concentration.
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Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (2) |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa. |
IHC-P
Use at an assay dependent concentration. |
Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
Target
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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
modificationsSubstrate 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
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Database links
- Entrez Gene: 387 Human
- Omim: 165390 Human
- SwissProt: P61586 Human
- Unigene: 247077 Human
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Alternative names
- Aplysia ras related homolog 12 antibody
- ARH12 antibody
- ARHA antibody
see all
Images
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RhoA antibody (ab54835) at 1ug/lane + HL-60 cell lysate (ab7914) at 25ug/lane.
This image was generated using the ascites version of the product.
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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
All lanes : Goat anti-mouse IgG
Predicted band size: 22 kDaThis image was generated using the ascites version of the product.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA antibody [1B12] (ab54835)
Paraffin embedded human lymphoma tissue stained for RhoA using ab54835 at 5 µg/ml in immunohistochemical analysis.
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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.
This image was generated using the ascites version of the product.
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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.
This image was generated using the ascites version of the product. -
ab54835 at 10 ug/ml staining RhoA in human Hela cells (ab150035) by Immunocytochemistry / Immunofluorescence.
This image was generated using the ascites version of the product.
Protocols
Datasheets and documents
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Datasheet download
References (77)
ab54835 has been referenced in 77 publications.
- Shao F et al. Oxycodone relieves permeability damage and apoptosis of oxygen-glucose deprivation/reoxygenation-induced brain microvascular endothelial cells through ras homolog family member A (RhoA)/ Rho-associated coiled-coil containing kinases (ROCK)/ myosin light chain 2 (MLC2) signal. Bioengineered 13:5205-5215 (2022). PubMed: 35170371
- Xia N et al. HNRNPC regulates RhoA to induce DNA damage repair and cancer-associated fibroblast activation causing radiation resistance in pancreatic cancer. J Cell Mol Med 26:2322-2336 (2022). PubMed: 35277915
- Hamuro J et al. Repressed miR-34a Expression Dictates the Cell Fate to Corneal Endothelium Failure. Invest Ophthalmol Vis Sci 63:22 (2022). PubMed: 35475886
- Nam S et al. RHOA protein expression correlates with clinical features in gastric cancer: a systematic review and meta-analysis. BMC Cancer 22:798 (2022). PubMed: 35854253
- Shi F et al. Relationship between the expression of ARHGAP25 and RhoA in non-small cell lung cancer and vasculogenic mimicry. BMC Pulm Med 22:377 (2022). PubMed: 36207695