Invitrogen

RhoA/RhoC Monoclonal Antibody (1B3-4A10)

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RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

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RhoA/RhoC Antibody (MA1-123) in ICC/IF

Immunofluorescence analysis of RhoA/RhoC was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RhoA/RhoC Mouse Monoclonal Antibody (Product # MA1-123) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius ove... View More

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF)

RhoA/RhoC Antibody in Immunohistochemistry (Paraffin) (IHC (P))

RhoA/RhoC Antibody in Immunohistochemistry (Paraffin) (IHC (P))

RhoA/RhoC Antibody in Immunohistochemistry (Paraffin) (IHC (P))

RhoA/RhoC Antibody in Western Blot (WB)

RhoA/RhoC Antibody in Western Blot (WB)

RhoA/RhoC Antibody in Western Blot (WB)

RhoA/RhoC Antibody in Immunoprecipitation (IP)

RhoA/RhoC Antibody

RhoA/RhoC Antibody

Product Image Gallery

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of RhoA/RhoC was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RhoA/RhoC Mouse Monoclonal Antibody (Product # MA1-123) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF) — Knockdown of RhoA/RhoC was achieved by transfecting HeLa cells with RhoA,RhoC specific siRNAs (Silencer® select Product # s758, s97). Immunofluorescence analysis was performed using untransfected HeLa cells (panels a, d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with RhoA and RhoC specific siRNAs (panel c,f). Cells were fixed, permeabilized, and probed with RhoA/RhoC Monoclonal Antibody (1B3-4A10) (Product # MA1-123, 5 µg/mL), followed by labelling with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor 488 (Product # A28175, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962) and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of specific cytoplasmic localization was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to RhoA/RhoC. The images were captured at 60X magnification.

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF) — 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 a Rho A/C monoclonal antibody (Product # MA1-123) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC 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.

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF) — 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 a Rho A/C monoclonal antibody (Product # MA1-123) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC 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.

RhoA/RhoC Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of RhoA/C (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 0.3%% BSA/TBST (Product # 37525) for 15 minutes at room temperature. Cells were probed with a RhoA/C monoclonal antibody (Product # MA1-123) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

RhoA/RhoC Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry 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 a Rho A/C monoclonal antibody (Product # MA1-123) 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.

RhoA/RhoC Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — 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 a Rho A/C monoclonal antibody (Product # MA1-123) 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.

RhoA/RhoC Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — 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 a Rho A/C monoclonal antibody (Product # MA1-123) 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.

RhoA/RhoC Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of SW 480 (Lane 1), SK-OV-3 (Lane 2), NIH/3T3 (Lane 3) and PC-12 (Lane 4). The blot was probed with Anti-RhoA/RhoC Monoclonal Antibody (Product # MA1-123, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 21 kDa band corresponding to RhoA/RhoC was observed across the cell lines tested.

RhoA/RhoC Antibody in Western Blot (WB) — Knockdown of RhoA/RhoC was achieved by transfecting SW-480 with RhoA and RhoC specific siRNAs (Silencer® select Product # s758, s97). Western blot analysis (Fig. a) was performed using membrane extracts from the RhoA and RhoC knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with RhoA/RhoC Monoclonal Antibody (1B3-4A10) (Product # MA1-123, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to RhoA/RhoC.

RhoA/RhoC Antibody in Western Blot (WB) — Western blot analysis of RhoA/C was performed by loading 25 µg of various whole cell lysates or 0.5 µg of purified protein (RhoA, RhoB, and a GST-RhoC fusion) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a RhoA/C monoclonal antibody (Product # MA1-123) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:15,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

RhoA/RhoC Antibody in Immunoprecipitation (IP) — Immunoprecipitation of RhoA/C was performed on HeLa cells. Antigen-antibody complexes were formed by incubating 500 µg of whole cell lysate with 2 µg of a RhoA/C monoclonal antibody (Product # MA1-123) overnight on a rocking platform at 4°C. The immune complexes were captured on 50 µL Protein A/G Plus Agarose (Product # 20423), washed extensively, and eluted with Lane Marker Reducing Sample Buffer (Product # 39000). HeLa cell lysate was run as a control (left lane). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a RhoA/C monoclonal antibody (Product # MA1-123) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with Clean-Blot IP Detection Reagent (Product # 21230) at a dilution of 1:2500 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

RhoA/RhoC Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with RhoA and RhoC siRNA and loss of signal was observed in Immunofluorescence using RhoA/RhoC Monoclonal Antibody (1B3-4A10) (Product # MA1-123). {KD}

RhoA/RhoC Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. SW-480 cells were transfected with RhoA and RhoC siRNA and loss of signal was observed in Western Blot using RhoA/RhoC Monoclonal Antibody (1B3-4A10) (Product # MA1-123). {KD}

产品信息

MA1-123

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:500-1:1,500

-

免疫组化（石蜡） (IHC (P))

1:50-1:500

-

免疫细胞化学 (ICC/IF)

5 µg/mL

-

免疫沉淀 (IP)

2 µg

-

产品规格

种属反应

Human, Mouse, Rat

宿主/亚型

Mouse / IgG

分类

Monoclonal

类型

Antibody

克隆号

1B3-4A10

抗原

Full-length human recombinant RhoA protein.

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偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

1 mg/mL

纯化类型

Protein A

保存液

PBS with 30% glycerol, 1mg/mL BSA

内含物

0.05% sodium azide

保存条件

-20°C

运输条件

Ambient (domestic); Wet ice (international)

AB_2536831

产品详细信息

MA1-123 detects GTPases RhoA and RhoC, but does not react with RhoB.

MA1-123 has been successfully used in Western blot, immunofluorescence, IHC (P) and immunoprecipitation applications with human and mouse samples.

靶标信息

RhoA is described as a small GTPase protein that regulates the actin cytoskeleton in the formation of stress fibers. RhoA is also included in the related proteins of the Ras superfamily, which involves proteins in the regulation and timing of cell division.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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引用该产品

生物信息学

蛋白别名: Aplysia ras-related homolog 12; aplysia ras-related homolog 9 (RhoC); aplysia ras-related homolog A; aplysia ras-related homolog A1; aplysia ras-related homolog A2; h12; h9; oncogene RHO H12; oncogene RHO H9; plysia ras-related homolog A2; Ras family member A; ras homolog 9 (RhoC); ras homolog A1; ras homolog A2; ras homolog gene family, member A; ras homolog gene family, member A1; ras homolog gene family, member A2; ras homolog gene family, member C; RAS-related homolog 9; Rho; Rho cDNA clone 12; Rho cDNA clone 9; Rho family GTPase; Rho-related GTP-binding protein RhoC; rhoC GTPase; SIG-61; Silica-induced gene 61 protein; small GTP binding protein RhoA; small GTP binding protein RhoC; Transforming protein RhoA

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基因别名: AI324259; ARH12; ARH9; ARHA; Arha1; Arha2; ARHC; H9; RHO12; RHOA; RHOC; RHOH12; RHOH9

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UniProt ID: (Human) P61586, (Human) P08134, (Rat) P61589, (Mouse) Q9QUI0, (Mouse) Q62159

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Entrez Gene ID: (Human) 387, (Human) 389, (Rat) 117273, (Mouse) 11848, (Mouse) 11853, (Rat) 295342

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GTPase activity protein binding GTP binding myosin binding signal transducer activity GDP binding protein domain specific binding Rho GDP-dissociation inhibitor binding nucleotide binding small GTPase

transforming growth factor beta receptor signaling pathway Rho protein signal transduction viral process cell migration substantia nigra development actin cytoskeleton organization platelet activation regulation of cell migration actin cytoskeleton reorganization positive regulation of cytokinesis regulation of actin cytoskeleton organization regulation of osteoblast proliferation Roundabout signaling pathway cleavage furrow formation apolipoprotein A-I-mediated signaling pathway positive regulation of NF-kappaB import into nucleus positive regulation of I-kappaB kinase/NF-kappaB signaling stress fiber assembly apical junction assembly endothelial cell migration ossification involved in bone maturation wound healing, spreading of cells positive regulation of neuron differentiation vascular endothelial growth factor receptor signaling pathway ephrin receptor signaling pathway phosphatidylinositol-mediated signaling negative regulation of axonogenesis positive regulation of axonogenesis negative chemotaxis regulation of small GTPase mediated signal transduction positive regulation of stress fiber assembly Wnt signaling pathway, planar cell polarity pathway positive regulation of lipase activity trabecula morphogenesis positive regulation of protein serine/threonine kinase activity negative regulation of cell migration involved in sprouting angiogenesis mitotic spindle assembly endothelial tube lumen extension skeletal muscle satellite cell migration mitotic cleavage furrow formation regulation of cell motility cytokinesis small GTPase mediated signal transduction cell morphogenesis response to hypoxia regulation of transcription from RNA polymerase II promoter cytoskeleton organization cell adhesion cell-matrix adhesion skeletal muscle tissue development regulation of actin polymerization or depolymerization metabolic process response to mechanical stimulus response to glucose regulation of neuron projection development cerebral cortex cell migration forebrain radial glial cell differentiation cell differentiation positive regulation of cell growth positive regulation of cell migration androgen receptor signaling pathway positive regulation of actin filament polymerization stress-activated protein kinase signaling cascade negative regulation of intracellular steroid hormone receptor signaling pathway response to drug negative regulation of I-kappaB kinase/NF-kappaB signaling response to amino acid positive regulation of cysteine-type endopeptidase activity involved in apoptotic process negative regulation of neuron apoptotic process positive regulation of neuron apoptotic process response to ethanol negative regulation of neuron differentiation positive regulation of translation positive regulation of cell adhesion positive regulation of vasoconstriction positive regulation of smooth muscle contraction neuron projection morphogenesis regulation of dendrite development response to glucocorticoid regulation of calcium ion transport negative regulation of cell death cellular response to progesterone stimulus positive regulation of podosome assembly regulation of neural precursor cell proliferation cell cycle integrin-mediated signaling pathway GTP metabolic process cell division signal transduction