- Datasheet
- Specific References (7)
- Protocols
Overview
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Product nameAnti-RhoA antibody
See all RhoA primary antibodies -
DescriptionRabbit polyclonal to RhoA
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, WBmore details
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Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Dog, Zebrafish, Orangutan
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human RhoA.
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Positive control
- This antibody gave a positive signal in the following whole cell lysates: HL60; MCF7; MDA-MB-231; HeLa; Human Platelets.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Assay kits
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Compatible Secondaries
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
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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.
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Sequence similaritiesBelongs to the small GTPase superfamily. Rho family.
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DomainThe basic-rich region is essential for yopT recognition and cleavage.
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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 localizationCell membrane. Cytoplasm > cytoskeleton.
- Information by UniProt
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Database links
- Entrez Gene: 395442 Chicken
- Entrez Gene: 338049 Cow
- Entrez Gene: 403954 Dog
- Entrez Gene: 387 Human
- Entrez Gene: 11848 Mouse
- Entrez Gene: 117273 Rat
- Omim: 165390 Human
- SwissProt: P61585 Cow
see all -
Alternative names
- Aplysia ras related homolog 12 antibody
- ARH12 antibody
- ARHA antibody
see all
Images
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Western blot - Anti-RhoA antibody (ab86297)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
All lanes : 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 -
Western blot - Anti-RhoA antibody (ab86297)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
All lanes : 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 -
Western blot - Anti-RhoA antibody (ab86297) -
Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody (ab86297)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
This product has been referenced in:
- Abou-Antoun TJ et al. Molecular and functional analysis of anchorage independent, treatment-evasive neuroblastoma tumorspheres with enhanced malignant properties: A possible explanation for radio-therapy resistance. PLoS One 13:e0189711 (2018). Read more (PubMed: 29298329) »
- Ye Y et al. Exosomal miR-141-3p regulates osteoblast activity to promote the osteoblastic metastasis of prostate cancer. Oncotarget 8:94834-94849 (2017). Read more (PubMed: 29212270) »
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