Name: Anti-RhoA antibody [EPR18134]
Brand: RabMAb
SKU: ab187027
Rating: 4 (1 reviews)

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Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18134] to RhoA
Suitable for: Flow Cyt (Intra), WB, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free PE

製品名

Anti-RhoA antibody [EPR18134]
RhoA 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR18134] to RhoA
由来種

Rabbit
アプリケーション

適用あり: Flow Cyt (Intra), WB, ICC/IFmore details
種交差性

交差種: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: Human RhoA full length protein; HeLa, HEK-293 C6, Raw264.7 and NIH/3T3 cell lysates, human fetal brain and fetal kidney lysates, mouse brain, kidney and spleen lysates, rat brain, kidney and spleen lysates. ICC/IF: Jurkat and K562 cells. Flow Cyt (intra): HeLa cells.
特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

製品の状態

Liquid
保存方法

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.
バッファー

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR18134
アイソタイプ

IgG
研究分野

Alternative Versions
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human RHOA knockout HEK-293T cell line (ab266592)
KO cell lysates
- Human RHOA knockout HEK-293T cell lysate (ab257637)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab187027の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
Flow Cyt (Intra)		1/200.
WB	(1)	1/5000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
ICC/IF		1/150.

特記事項
Flow Cyt (Intra) 1/200.
WB 1/5000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
ICC/IF 1/150.

機能

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.
配列類似性

Belongs to the small GTPase superfamily. Rho family.
ドメイン

The basic-rich region is essential for yopT recognition and cleavage.
翻訳後修飾

Substrate 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.
細胞内局在

Cell membrane. Cytoplasm > cytoskeleton.
Target information above from: UniProt accession P61586 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 387 Human
- Entrez Gene: 11848 Mouse
- Entrez Gene: 117273 Rat
- Omim: 165390 Human
- SwissProt: P61586 Human
- SwissProt: Q9QUI0 Mouse
- SwissProt: P61589 Rat
- Unigene: 247077 Human
- Unigene: 757 Mouse
- Unigene: 107401 Rat
see all
別名
- Aplysia ras related homolog 12 antibody
- ARH12 antibody
- ARHA antibody
- H 12 antibody
- H12 antibody
- Oncogene RHO H12 antibody
- Ras homolog family member A antibody
- Ras homolog gene family member A antibody
- Rho A antibody
- Rho cDNA clone 12 antibody
- RHO H12 antibody
- RHO12 antibody
- RHOA antibody
- RHOA_HUMAN antibody
- RHOH12 antibody
- Small GTP binding protein Rho A antibody
- Transforming protein Rho A antibody
- Transforming protein RhoA antibody
see all

Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

Lanes 1 & 3 : Wild-type HEK-293T cell lysate
Lanes 2 & 4 : RHOA knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 22 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Lanes 1- 4: Merged signal (red and green). Green - ab187027 observed at 21 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab187027 was shown to react with RhoA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266592 (knockout cell lysate ab257637) was used. Wild-type HEK-293T and RHOA knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187027 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] (ab187027)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling RhoA with ab187027 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Jurkat cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
1. ab187027 at 1/150 dilution followed by ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/1000 dilution

Lane 1 : Wild-type Hek293T whole cell lysate
Lane 2 : RHOA knockout Hek293T whole cell lysate
Lane 3 : Jurkat whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 22 kDa

Lanes 1 - 3: Merged signal (red and green). Green - ab187027 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab187027 was shown to specifically react with RhoA in wild-type Hek293T cells as signal was lost in RHOA knockout cells. Wild-type and RHOA knockout samples were subjected to SDS-PAGE. Ab187027 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

Lane 1 : Human RhoA full length protein
Lane 2 : Human RhoB full length protein
Lane 3 : Human RhoC full length protein

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 1 second

5% NFDM/TBST: Blocking and diluting buffer.

Human RhoA full length protein (ab101594) contains aa1-193 with His-Tag^® at N-Terminus; Human RhoB full length protein (ab107139) contains aa1-193 with His-Tag^® at N-Terminus; Human RhoC full length protein (ab98085) contains aa1-190 with His-Tag^® at N-Terminus.
Flow Cytometry (Intracellular) - Anti-RhoA antibody [EPR18134] (ab187027)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling RhoA with ab187027 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and a unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 3 minutes

5% NFDM/TBST: Blocking and diluting buffer.
Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 3 minutes

5% NFDM/TBST: Blocking and diluting buffer.
Western blot - Anti-RhoA antibody [EPR18134] (ab187027)

All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat kidney lysate
Lane 6 : Rat spleen lysate
Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
Lane 8 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 3 minutes

5% NFDM/TBST: Blocking and diluting buffer.
Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] (ab187027)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling RhoA with ab187027 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
1. ab187027 at 1/150 dilution followed by ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Anti-RhoA antibody [EPR18134] (ab187027)

Flow cytometry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab187027 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab187027 は 95 報の論文で使用されています。

Qin X et al. LAMB3 Promotes Myofibrogenesis and Cytoskeletal Reorganization in Endometrial Stromal Cells via the RhoA/ROCK1/MYL9 Pathway. Cell Biochem Biophys 82:127-137 (2024). PubMed: 37801199
Lin X et al. Targeting of G-protein coupled receptor 40 alleviates airway hyperresponsiveness through RhoA/ROCK1 signaling pathway in obese asthmatic mice. Respir Res 24:56 (2023). PubMed: 36803977
Huang S et al. Corydalis decumbens Alleviates the Migration, Phagocytosis, and Inflammatory Response of Macrophages. Evid Based Complement Alternat Med 2023:7000477 (2023). PubMed: 36874618
Wang J et al. RhoA promotes osteoclastogenesis and regulates bone remodeling through mTOR-NFATc1 signaling. Mol Med 29:49 (2023). PubMed: 37020186
Tang X et al. Exploiting synergistic effect of CO/NO gases for soft tissue transplantation using a hydrogel patch. Nat Commun 14:2417 (2023). PubMed: 37105981

View all Publications for this product

レビューと Q&A

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レビューを送る質問を送る

Application

Western blot

Sample

Human Cell lysate - whole cell (Human Cancer Cell line)

Gel Running Conditions

Reduced Denaturing (12)

Loading amount

10 µg

Specification

Human Cancer Cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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投稿 Nov 12 2021

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Key features and details

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製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

KO cell lines

KO cell lysates

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

配列類似性

ドメイン

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

Certificate of Compliance

参考文献 (95)

レビューと Q&A

Western blot abreview for Anti-RhoA antibody [EPR18134]