Anti-ROCK1 抗体 [EPR638Y] (ab134181)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR638Y] to ROCK1
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-ROCK1 antibody [EPR638Y]
ROCK1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR638Y] to ROCK1 -
由来種
Rabbit -
特異性
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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アプリケーション
適用あり: Flow Cyt (Intra), WB, IP, IHC-Pmore 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.
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ポジティブ・コントロール
- WB: HeLa, HAP1, and HEK293 cell lysates, and Mouse brain and Rat brain tissue lysates IHC: Human testis tissue and Human breast carcinoma tissue IP: HeLa cell lysate Flow Cyt (Intra): HeLa cells
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR638Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Corresponding Antibody
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Isotype control
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Recombinant Protein
アプリケーション
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ab134181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 |
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Flow Cyt (Intra) |
1/150.
Not suitable for unpurified form. |
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WB | (1) |
1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa).
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IP |
1/60 - 1/120.
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IHC-P |
1/1200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols For unpurified use at 1/50 - 1/100. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
特記事項 |
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Flow Cyt (Intra)
1/150. Not suitable for unpurified form. |
WB
1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa). |
IP
1/60 - 1/120. |
IHC-P
1/1200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols For unpurified use at 1/50 - 1/100. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
ターゲット情報
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機能
Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of DAPK3, GFAP, LIMK1, LIMK2, MYL9/MLC2, PFN1 and PPP1R12A. Phosphorylates FHOD1 and acts synergistically with it to promote SRC-dependent non-apoptotic plasma membrane blebbing. Phosphorylates JIP3 and regulates the recruitment of JNK to JIP3 upon UVB-induced stress. Acts as a suppressor of inflammatory cell migration by regulating PTEN phosphorylation and stability. Acts as a negative regulator of VEGF-induced angiogenic endothelial cell activation. Required for centrosome positioning and centrosome-dependent exit from mitosis. Plays a role in terminal erythroid differentiation. May regulate closure of the eyelids and ventral body wall by inducing the assembly of actomyosin bundles. Promotes keratinocyte terminal differentiation. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization. -
組織特異性
Detected in blood platelets. -
配列類似性
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 PH domain.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain.
Contains 1 REM (Hr1) repeat. -
ドメイン
The C-terminal auto-inhibitory domain interferes with kinase activity. RHOA binding leads to a conformation change and activation of the kinase. Truncated ROCK1 is constitutively activated. -
翻訳後修飾
Autophosphorylated on serine and threonine residues.
Cleaved by caspase-3 during apoptosis. This leads to constitutive activation of the kinase and membrane blebbing. -
細胞内局在
Cytoplasm. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole. Golgi apparatus membrane. Cell projection, bleb. Cytoplasm, cytoskeleton. Cell membrane. Cell projection, lamellipodium. Cell projection, ruffle. Associated with the mother centriole and an intercentriolar linker. Colocalizes with ITGB1BP1 and ITGB1 at the cell membrane predominantly in lamellipodia and membrane ruffles, but also in retraction fibers. Localizes at the cell membrane in an ITGB1BP1-dependent manner (By similarity). A small proportion is associated with Golgi membranes. - Information by UniProt
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参照データベース
- Entrez Gene: 6093 Human
- Entrez Gene: 19877 Mouse
- Entrez Gene: 81762 Rat
- Omim: 601702 Human
- SwissProt: Q13464 Human
- SwissProt: P70335 Mouse
- SwissProt: Q63644 Rat
- Unigene: 306307 Human
see all -
別名
- coiled-coil-containing protein kinase 1 antibody
- coiled-coil-containing protein kinase I antibody
- MGC131603 antibody
see all
画像
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Lane 1: Wild type HAP1 whole cell lysate (40 µg)
Lane 2: ROCK1 knockout HAP1 whole cell lysate (40 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab134181 observed at 165 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab134181 was shown to specifically react with ROCK1 when ROCK1 knockout samples were used. Wild-type and ROCK1 knockout samples were subjected to SDS-PAGE. Ab134181 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling ROCK1 with Purified ab134181 at 1:1200 (1.28 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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All lanes : Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : Mouse brain
Lane 3 : Rat brain
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 158 kDa
Observed band size: 158 kDa -
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling ROCK1 with purified ab134181 at 1/150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150081) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue sections labeling ROCK1 with Purified ab134181 at 1:1200 (1.28 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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All lanes : Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ROCK1 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 158 kDa
Observed band size: 160 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab134181 observed at 160 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab134181 was shown to react with ROCK1 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264780 (CRISPR/Cas9 edited cell lysate ab257642) lane below 160kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ROCK1 CRISPR/Cas9 edited HeLa 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. ab134181 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 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.
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Purified ab134181 at 1/120 dilution (2µg) immunoprecipitating ROCK1 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab134181 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134181 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 158 kDa
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (34)
ab134181 は 34 報の論文で使用されています。
- Li C et al. Rho kinase inhibition ameliorates vascular remodeling and blood pressure elevations in a rat model of apatinib-induced hypertension. J Hypertens 40:675-684 (2022). PubMed: 34862331
- Pan ZN et al. LRRK2 regulates actin assembly for spindle migration and mitochondrial function in mouse oocyte meiosis. J Mol Cell Biol 14:N/A (2022). PubMed: 34918122
- Li X et al. Erianin Controls Collagen-Mediated Retinal Angiogenesis via the RhoA/ROCK1 Signaling Pathway Induced by the alpha2/beta1 Integrin-Collagen Interaction. Invest Ophthalmol Vis Sci 63:27 (2022). PubMed: 35060996
- Chen R et al. CID16020046, a GPR55 antagonist, attenuates sepsis‑induced acute kidney injury. Mol Med Rep 25:N/A (2022). PubMed: 35244189
- Wang H et al. Circ_0000064 promotes high glucose-induced renal tubular epithelial cells injury to facilitate diabetic nephropathy progression through miR-532-3p/ROCK1 axis. BMC Endocr Disord 22:67 (2022). PubMed: 35291991