Anti-eNOS 抗体 [M221] (ab76198)
Key features and details
- Mouse monoclonal [M221] to eNOS
- Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)
- Reacts with: Mouse, Human
- Isotype: IgG1
製品の概要
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製品名
Anti-eNOS antibody [M221]
eNOS 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [M221] to eNOS -
由来種
Mouse -
特異性
ab76198 is not predicted to react with other NOS family members due to low homology.
This antibody detects eNOS in mouse and but at a lower intensity than in human. If you are working in mouse, we would recommend using no more than 1% milk as the blocking agent for optimal signal strength.
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アプリケーション
適用あり: IHC-P, ICC/IF, WB, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, Human -
免疫原
Recombinant fragment corresponding to Mouse eNOS aa 1024-1202 (C terminal).
Database link: P70313 -
ポジティブ・コントロール
- Human umbilical vein endothelial cells untreated and treated with lambda phosphatase. Mouse placenta lysate. Huvec lysate. IHC-P: FFPE human normal placenta tissue sections.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 -20°C. Stable for 12 months at -20°C. -
バッファー
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 50% Glycerol, PBS -
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
M221 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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ab76198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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IHC-P |
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (1) |
Use at an assay dependent concentration.
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WB | (11) |
1/500 - 1/1000. Predicted molecular weight: 133 kDa.
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Flow Cyt (Intra) |
Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
特記事項 |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
WB
1/500 - 1/1000. Predicted molecular weight: 133 kDa. |
Flow Cyt (Intra)
Use 2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets.
Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1. -
組織特異性
Platelets, placenta, liver and kidney. -
関連疾患
Variation in NOS3 seem to be associated with susceptibility to coronary spasm. -
配列類似性
Belongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. -
翻訳後修飾
Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity (By similarity). Phosphorylation of Ser-114 by CDK5 reduces activity. -
細胞内局在
Cell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle and which is favored by interaction with NOSIP and results in a reduced enzymatic activity. - Information by UniProt
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参照データベース
- Entrez Gene: 4846 Human
- Entrez Gene: 18127 Mouse
- Omim: 163729 Human
- SwissProt: P29474 Human
- SwissProt: P70313 Mouse
- Unigene: 647092 Human
- Unigene: 258415 Mouse
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別名
- cNOS antibody
- Constitutive NOS antibody
- EC NOS antibody
see all
画像
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All lanes : Anti-eNOS antibody [M221] (ab76198) at 1/500 dilution
Lane 1 : Huvec cell lysates
Lane 2 : Mouse placenta lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti Mouse IR680 at 1/10000 dilution
Predicted band size: 133 kDaThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab76198 overnight at 4°C. Antibody binding was detected using Goat anti Mouse IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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IHC image of eNOS staining in human normal placenta*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab76198, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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ab76198 staining eNOS in human umbilical vein endothelial cells. Cells with fixed with paraformaldehyde.
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Overlay histogram showing HEK293 cells stained with ab76198 (red line). The cells were fixed with 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 (ab76198, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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All lanes : Anti-eNOS antibody [M221] (ab76198) at 1/1000 dilution
Lane 1 : human umbilical vein endothelial
cells, untreated
Lane 2 : human umbilical vein endothelial
cells, treated with lambda phosphatase
Predicted band size: 133 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
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 (143)
ab76198 は 143 報の論文で使用されています。
- Su Y et al. Blending with transition metals improves bioresorbable zinc as better medical implants. Bioact Mater 20:243-258 (2023). PubMed: 35702610
- Li T et al. Intraoperative Hypothermia Induces Vascular Dysfunction in the CA1 Region of Rat Hippocampus. Brain Sci 12:N/A (2022). PubMed: 35741578
- Luo C et al. NLRP3 downregulation enhances engraftment and functionality of adipose-derived stem cells to alleviate erectile dysfunction in diabetic rats. Front Endocrinol (Lausanne) 13:913296 (2022). PubMed: 35937790
- Chen T et al. LncRNA Airn maintains LSEC differentiation to alleviate liver fibrosis via the KLF2-eNOS-sGC pathway. BMC Med 20:335 (2022). PubMed: 36171606
- Li Z et al. Cholesterol efflux regulator ABCA1 exerts protective role against high shear stress-induced injury of HBMECs via regulating PI3K/Akt/eNOS signaling. BMC Neurosci 23:61 (2022). PubMed: 36335301