Anti-RhoA + C antibody [1B34A10] (ab175359)

Mouse Monoclonal RHOA antibody. Suitable for IP, WB, IHC-P, ICC/IF and reacts with Human, Mouse samples. Immunogen corresponding to Recombinant Full Length Protein corresponding to Human RHOA.

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Images

Western blot - Anti-RhoA + C antibody [1B34A10] (AB175359), expandable thumbnail

Key facts

Isotype: IgG
Host species: Mouse
Storage buffer: Preservative: 0.05% Sodium azide
Constituents: 69% PBS, 30% Glycerol (glycerin, glycerine), 0.1% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

Recombinant Full Length Protein corresponding to Human RHOA. Database link P61586

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	IHC-P	ICC/IF
Human	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Use 2μg.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500 - 1/1500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100 - 1/200	Notes -
Species Human	Dilution info 1/100 - 1/200	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/200	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Target data

See full target information RHOA

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 as cytoskeletal dynamics, cell migration and cell cycle (PubMed:23871831). Regulates a signal transduction pathway linking plasma membrane receptors to the assembly of focal adhesions and actin stress fibers (PubMed:31570889, PubMed:8910519, PubMed:9121475). Involved in a microtubule-dependent signal that is required for the myosin contractile ring formation during cell cycle cytokinesis (PubMed:12900402, PubMed:16236794). Plays an essential role in cleavage furrow formation. Required for the apical junction formation of keratinocyte cell-cell adhesion (PubMed:20974804, PubMed:23940119). Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly (PubMed:19934221). 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 (PubMed:20937854). Regulates KCNA2 potassium channel activity by reducing its location at the cell surface in response to CHRM1 activation; promotes KCNA2 endocytosis (PubMed:19403695, PubMed:9635436). Acts as an allosteric activator of guanine nucleotide exchange factor ECT2 by binding in its activated GTP-bound form to the PH domain of ECT2 which stimulates the release of PH inhibition and promotes the binding of substrate RHOA to the ECT2 catalytic center (PubMed:31888991). May be an activator of PLCE1 (PubMed:16103226). In neurons, involved in the inhibition 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). Acts as a regulator of platelet alpha-granule release during activation and aggregation of platelets (By similarity). When activated by DAAM1 may signal centrosome maturation and chromosomal segregation during cell division. May also be involved in contractile ring formation during cytokinesis. (Microbial infection) Serves as a target for the yopT cysteine peptidase from Yersinia pestis, vector of the plague.

Additional Targets

RHOC

Alternative names

ARH12, ARHA, RHO12, RHOA, Transforming protein RhoA, Rho cDNA clone 12, h12

Recommended products

Mouse Monoclonal RHOA antibody. Suitable for IP, WB, IHC-P, ICC/IF and reacts with Human, Mouse samples. Immunogen corresponding to Recombinant Full Length Protein corresponding to Human RHOA.

Key facts

Isotype: IgG
Host species: Mouse
Storage buffer: Preservative: 0.05% Sodium azide
Constituents: 69% PBS, 30% Glycerol (glycerin, glycerine), 0.1% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: Recombinant Full Length Protein corresponding to Human RHOA. Database link P61586
Clone number: 1B34A10
Purification technique: Affinity purification Protein A
Specificity: ab175359 detects RhoA and RhoC, but does not react with RhoB.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RhoA also known as Ras homolog family member A is a small GTPase with a molecular mass of approximately 21 kDa. This protein belongs to the Rho family of GTPases and is widespread in various tissues particularly in the brain and heart. RhoA acts as a molecular switch cycling between an active GTP-bound state and an inactive GDP-bound state. Its primary function is to regulate the actin cytoskeleton influencing cell shape motility and division. It localizes mainly in the cytoplasm and occasionally associates with the plasma membrane.

Biological function summary

RhoA plays a role in several cellular processes such as morphogenesis migration and contraction by forming complexes notably with Rho-associated protein kinases (ROCK1 and ROCK2). Through these interactions RhoA controls cytoskeletal dynamics enabling cells to respond to extracellular signals. Additionally RhoA has a role in maintaining cell polarity and adhesion important for tissue organization and growth. Its regulation is significant in various cell types ensuring proper cellular responses during physiological activities.

Pathways

Four words immediately come to mind signal transduction pathways are where RhoA functions prominently especially the Rho/ROCK pathway. This pathway influences the rearrangement of the actin cytoskeleton and regulates cell proliferation and survival. RhoA also participates in the Wnt signaling pathway interacting with proteins such as β-catenin to affect gene expression. These pathways illustrate RhoA's role in integrating extracellular cues to elicit appropriate cellular outcomes linking it to other Rho family members like Rac1 and Cdc42 which have overlapping yet distinct functions.

Associated diseases and disorders

Four words are readily apparent aberrant RhoA signaling associates with various pathologies notably cancer and cardiovascular diseases. In cancer dysregulation of RhoA leads to increased cell migration and invasion contributing to metastasis. RhoA's link to cardiovascular diseases includes the regulation of vascular smooth muscle contraction impacting blood pressure and heart function. The RhoA-related ROCK proteins play an important role in these diseases as their increased activity can exacerbate pathological conditions demonstrating RhoA's potential as a therapeutic target.

Product promise

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9 product images

Western blot - Anti-RhoA + C antibody [1B34A10] (ab175359)
All lanes: Western blot - Anti-RhoA + C antibody [1B34A10] (ab175359) at 1/1000 dilution
Lane 1: SW 480 whole cell extract at 30 µg
Lane 2: SK-OV-3 whole cell extract at 30 µg
Lane 3: NIH-3T3 whole cell extract at 30 µg
Lane 4: PC-12 whole cell extract at 30 µg
Secondary
All lanes: Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/4000 dilution
Predicted band size: 22 kDa
Immunocytochemistry/ Immunofluorescence - Anti-RhoA + C antibody [1B34A10] (ab175359)
Immunofluorescent analysis of Rho A/C (green) showing staining in the in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab175359 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
Immunocytochemistry/ Immunofluorescence - Anti-RhoA + C antibody [1B34A10] (ab175359)
Immunofluorescent analysis of Rho A/C (green) showing staining in the in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab175359 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + C antibody [1B34A10] (ab175359)
Immunohistochemistry analysis of Rho A/C showing staining in the cytoplasm of paraffin-embedded human hepatocarcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab175359 diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + C antibody [1B34A10] (ab175359)
Immunohistochemistry analysis of Rho A/C showing staining in the cytoplasm and membrane of paraffin-embedded mouse liver tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab175359 diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RhoA + C antibody [1B34A10] (ab175359)
mmunohistochemistry analysis of Rho A/C showing staining in the cytoplasm and membrane of paraffin-embedded human prostate carcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab175359 diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Immunoprecipitation - Anti-RhoA + C antibody [1B34A10] (ab175359)
Immunoprecipitation analysis of RhoA + C was performed on HeLa cells. Antigen-antibody complexes were formed by incubating 500μg of whole cell lysate with 2μg ab175359 (lane 2).
HeLa cell lysate was run as a control (lane 1).
For WB detection, ab175359 was used at 1/1000 dilution.
All lanes: Immunoprecipitation - Anti-RhoA + C antibody [1B34A10] (ab175359)
Developed using the ECL technique.
Predicted band size: 22 kDa
Western blot - Anti-RhoA + C antibody [1B34A10] (ab175359)
All lanes: Western blot - Anti-RhoA + C antibody [1B34A10] (ab175359) at 1/1000 dilution
Lane 1: HeLa whole cell lysate at 25 µg
Lane 2: A431 whole cell lysate at 25 µg
Lane 3: HEK293T whole cell lysate at 25 µg
Lane 4: U2 OS whole cell lysate at 25 µg
Lane 5: NIH 3T3 whole cell lysate at 25 µg
Lane 6: K562 whole cell lysate at 25 µg
Lane 7: RhoA purified protein at 0.5 µg
Lane 8: RhoB purified protein at 0.5 µg
Lane 9: RhoC purified protein with proprietary tag at 0.5 µg
Secondary
All lanes: goat anti-mouse IgG-HRP at 1/15000 dilution
Developed using the ECL technique.
Predicted band size: 22 kDa
Immunocytochemistry/ Immunofluorescence - Anti-RhoA + C antibody [1B34A10] (ab175359)
Immunofluorescent analysis of HeLa cells (formalin-fixed, 0.1% Triton X-100 permeabilized) labeling RhoA + C with ab175359 at 1/100 dilution followed with DyLight 488 goat anti-mouse IgG secondary antibody at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.