Product: Phospho-RhoA (Ser188) Antibody
Catalog: AF8020
Description: Rabbit polyclonal antibody to Phospho-RhoA (Ser188)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Sheep, Dog
Mol.Wt.: 22kDa; 22kD(Calculated).
Uniprot: P61586
RRID: AB_2840083

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 100ul $350 In stock
 200ul $450 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Sheep(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-RhoA (Ser188) Antibody detects endogenous levels of RhoA only when phosphorylated at Ser188.
RRID:
AB_2840083
Cite Format: Affinity Biosciences Cat# AF8020, RRID:AB_2840083.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Aplysia ras related homolog 12; ARH12; ARHA; H 12; H12; Oncogene RHO H12; Ras homolog family member A; Ras homolog gene family member A; Rho A; Rho cDNA clone 12; RHO H12; RHO12; RHOA; RHOA_HUMAN; RHOH12; Small GTP binding protein Rho A; Transforming protein Rho A; Transforming protein RhoA;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Sequence:
MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATRAALQARRGKKKSGCLVL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Xenopus
63
Horse
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P61586 As Substrate

Site PTM Type Enzyme
K7 Ubiquitination
T19 Phosphorylation
S26 Phosphorylation
K27 Sumoylation
Y34 Phosphorylation
Y42 Phosphorylation
T60 Phosphorylation
Y66 Phosphorylation
S88 Phosphorylation P27361 (MAPK3)
T100 Phosphorylation P27361 (MAPK3)
K104 Ubiquitination
C107 S-Nitrosylation
K118 Ubiquitination
K119 Methylation
K119 Ubiquitination
T127 Phosphorylation
R128 Methylation
K133 Ubiquitination
K135 Ubiquitination
K140 Ubiquitination
Y156 Phosphorylation
C159 S-Nitrosylation
S160 Phosphorylation
K164 Ubiquitination
S188 Phosphorylation Q9H2G2 (SLK) , P17612 (PRKACA) , Q13976 (PRKG1)

Research Backgrounds

Function:

Small GTPase which cycles between an active GTP-bound and an inactive GDP-bound state. Mainly associated with cytoskeleton organization, in active state binds to a variety of effector proteins to regulate cellular responses such cytoskeletal dynamics, cell migration and cell cycle. 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. 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 KCNA2 potassium channel activity by reducing its location at the cell surface in response to CHRM1 activation; promotes KCNA2 endocytosis. May be an activator of PLCE1. In neurons, involved in the inhibiton of the initial spine growth. Upon activation by CaMKII, modulates dendritic spine structural plasticity by relaying CaMKII transient activation to synapse-specific, long-term signaling (By similarity).

(Microbial infection) Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague.

PTMs:

(Microbial infection) Substrate for botulinum ADP-ribosyltransferase.

(Microbial infection) 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.

(Microbial infection) 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.

(Microbial infection) Glycosylated at Tyr-34 by Photorhabdus asymbiotica toxin PAU_02230. Mono-O-GlcNAcylation by PAU_02230 inhibits downstream signaling by an impaired interaction with diverse regulator and effector proteins of Rho and leads to actin disassembly.

Phosphorylation by PRKG1 at Ser-188 inactivates RHOA signaling. Phosphorylation by SLK at Ser-188 in response to AGTR2 activation (By similarity).

Ubiquitinated by the BCR(KCTD13) and BCR(TNFAIP1) E3 ubiquitin ligase complexes, leading to its degradation by the proteasome, thereby regulating the actin cytoskeleton and synaptic transmission in neurons.

Subcellular Location:

Cell membrane>Lipid-anchor>Cytoplasmic side. Cytoplasm>Cytoskeleton. Cleavage furrow. Cytoplasm>Cell cortex. Midbody. Cell projection>Lamellipodium. Cell projection>Dendrite.
Note: Localized to cell-cell contacts in calcium-treated keratinocytes (By similarity). Translocates to the equatorial region before furrow formation in a ECT2-dependent manner. Localizes to the equatorial cell cortex (at the site of the presumptive furrow) in early anaphase in an activated form and in a myosin- and actin-independent manner.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

Interacts with ARHGEF28 (By similarity). Interacts (via GTP-bound form) with RIPOR1 (via N-terminus); this interaction links RHOA to STK24 and STK26 kinases. Interacts with RIPOR2 (via active GTP- or inactive GDP-bound forms) isoform 1 and isoform 2; these interactions are direct, block the loading of GTP to RHOA and decrease upon chemokine CCL19 stimulation in primary T lymphocytes. Binds PRKCL1, ROCK1 and ROCK2. Interacts with ARHGEF2, ARHGEF3, NET1 and RTKN. Interacts with PLCE1 and AKAP13. Interacts with DIAPH1. Interacts (in the constitutively activated, GTP-bound form) with DGKQ. Interacts with RACK1; enhances RHOA activation. Interacts with PKP4; the interaction is detected at the midbody. Interacts (GTP-bound form preferentially) with PKN2; the interaction stimulates autophosphorylation and phosphorylation of PKN2. Interacts with ARHGDIA; this interaction inactivates and stabilizes RHOA. Interacts with ARHGDIB. Interacts (GTP-bound form) with KCNA2 (via cytoplasmic N-terminal domain).

(Microbial infection) Interacts with yopT from Yersinia pestis.

(Microbial infection) Interacts with human respiratory syncytial virus (HRSV) protein F; this interaction facilitates virus-induced syncytium formation.

Family&Domains:

The basic-rich region is essential for yopT recognition and cleavage.

Belongs to the small GTPase superfamily. Rho family.

Research Fields

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Adherens junction.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Wnt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Bacterial > Bacterial invasion of epithelial cells.

· Human Diseases > Infectious diseases: Bacterial > Pathogenic Escherichia coli infection.

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Circulatory system > Vascular smooth muscle contraction.   (View pathway)

· Organismal Systems > Development > Axon guidance.   (View pathway)

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > T cell receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Digestive system > Pancreatic secretion.

References

1). Comparative Phosphoproteomics of Neuro-2a Cells under Insulin Resistance Reveals New Molecular Signatures of Alzheimer’s Disease. International Journal of Molecular Sciences, 2022 (PubMed: 35055191) [IF=5.6]

Application: WB    Species: Mice    Sample: brain tissues

Figure 5 Comparative informatic analysis of phosphoproteomes of two different insulin-resistant conditions. (A) Comparative canonical pathway analysis between the phosphoproteomes. The positive and negative z-scores are shown in orange and blue, respectively. “Pal” and “TNF” indicate palmitate-induced insulin-resistant and TNF-α-induced insulin-resistant conditions, respectively. (B) The integrin pathway was the most downregulated pathway between the phosphoproteomes. The increased and decreased phosphorylation levels are shown in red and green, respectively. The red circles indicate the phosphoproteins commonly regulated by different insulin-resistant conditions. (C) Western blot of phospho-RhoA under palmitate-induced insulin-resistant conditions. Conditioned cell lysates were electrophoresed and blotted. “Insulin” indicates 100 nM of insulin treatment for 10 min. (D) The adenosine monophosphate-activated protein kinase pathway was the most downregulated pathway between the phosphoproteomes. (E) Western blot of phospho-protein kinase B and phospho-acetyl-CoA carboxylase under different insulin-resistant conditions. Conditioned cell lysates were electrophoresed and blotted. “Insulin” indicates 100 nM of insulin treatment for 10 min. (F) Comparative analysis for disease relation and biological functions. The positive and negative z-scores are shown in orange and blue color, respectively. “Pal” and “TNF” indicate palmitate-induced insulin-resistant condition and TNF-α-induced insulin-resistant condition, respectively.

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