Anti-ROCK2 + ROCK1 抗体 [EP786Y] (ab45171)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1
- Suitable for: Flow Cyt (Intra), IHC-Fr, WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Cow, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-ROCK2 + ROCK1 antibody [EP786Y] -
製品の詳細
Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1 -
由来種
Rabbit -
特異性
This antibody recognizes both the cleaved C-terminus of ROCK 1 (30 kDa) and full length protein (158 kDa). The immunogen used for this product shares 83% homology with ROCK2 and has been shown to bind recombinant human ROCK2, please see western blot images below.
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アプリケーション
適用あり: Flow Cyt (Intra), IHC-Fr, WB, IHC-P, ICC/IF, IPmore details -
種交差性
交差種: Mouse, Rat, Cow, Human -
免疫原
Synthetic peptide within Human ROCK1 aa 1100-1200 (C terminal). The exact sequence is proprietary.
Database link: Q13464 -
ポジティブ・コントロール
- WB: Untreated, Calyculin A treated and Camptothecin treated HeLa whole cell lysate (ab150035). Jurkat, Ramos, PC-12 and RAW264.7 cell lysates. IHC-P: Human adenocarcinoma of the colon and thyroid gland carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate (ab150035).
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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. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 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
EP786Y -
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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Conjugation kits
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Isotype control
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Positive Controls
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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ab45171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) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-Fr |
Use at an assay dependent concentration. PubMed: 19295659
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WB | (4) |
1/2000 - 1/10000. Predicted molecular weight: 158 kDa.
For unpurified use at 1/500. |
IHC-P | (2) |
1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/50 - 1/500.
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IP |
1/40 - 1/50.
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特記事項 |
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Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-Fr
Use at an assay dependent concentration. PubMed: 19295659 |
WB
1/2000 - 1/10000. Predicted molecular weight: 158 kDa. For unpurified use at 1/500. |
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/50 - 1/500. |
IP
1/40 - 1/50. |
ターゲット情報
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細胞内局在
ROCK2: Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane. ROCK1: 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. -
参照データベース
- Entrez Gene: 282041 Cow
- Entrez Gene: 785911 Cow
- Entrez Gene: 6093 Human
- Entrez Gene: 9475 Human
- Entrez Gene: 19877 Mouse
- Entrez Gene: 19878 Mouse
- Entrez Gene: 25537 Rat
- Entrez Gene: 81762 Rat
see all
画像
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All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/5000 dilution (purified)
Lane 1 : HeLa cell lysate - treated with Calyculin A
Lane 2 : HeLa cell lysate - treated with Camptothecin
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : Ramos cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 158 kDa
Observed band size: 158 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling ROCK2 + ROCK1 with purified ab45171 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ROCK2 + ROCK1 with purified ab45171 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150078, an Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/50) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).
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Overlay histogram showing HeLa cells stained with unpurified ab45171 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45171, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/1000 dilution
Lane 1 : Recombinant Human ROCK1 protein (aa 1114 to 1354) (30 kDa)
Lane 2 : Recombinant Human ROCK2 protein (aa 1132 to 1388) (30 kDa)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 158 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?Loading control: Anti-6X His tag® antibody [EPR20547] (ab213204)
Blocking buffer and concentration: 5% NFDM/TBST
Exposure Times:
Lane 1: 3 seconds
Lane 2: 7 seconds
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ab45171 (purified) at 1/40 immunoprecipitating ROCK2 + ROCK1 in HeLa cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/5000 dilution (purified)
Lane 1 : PC-12 cell lysate
Lane 2 : RAW264.7 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 158 kDa
Observed band size: 158 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling ROCK2 + ROCK1 with unpurified ab45171.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
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 (126)
ab45171 は 126 報の論文で使用されています。
- Wang X et al. Yes-associated protein reacts differently in vascular smooth muscle cells under different intensities of mechanical stretch. Aging (Albany NY) 14:286-296 (2022). PubMed: 34983026
- Wu D et al. CTNNAL1 participates in the regulation of mucus overproduction in HDM-induced asthma mouse model through the YAP-ROCK2 pathway. J Cell Mol Med 26:1656-1671 (2022). PubMed: 35092120
- Huang S et al. Cytoplasmic Clusterin Suppresses Lung Cancer Metastasis by Inhibiting the ROCK1-ERK Axis. Cancers (Basel) 14:N/A (2022). PubMed: 35626071
- Li J et al. WJ-MSCs intervention may relieve intrauterine adhesions in female rats via TGF-β1-mediated Rho/ROCK signaling inhibition. Mol Med Rep 23:N/A (2021). PubMed: 33179074
- Zou YJ et al. RAB14 GTPase is essential for actin-based asymmetric division during mouse oocyte maturation. Cell Prolif 54:e13104 (2021). PubMed: 34323331