Anti-Smad4 抗体 [EP618Y] (ab40759)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP618Y] to Smad4
- Suitable for: WB, 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-Smad4 antibody [EP618Y]
Smad4 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP618Y] to Smad4 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-Pmore details
適用なし: Flow Cyt,ICC/IF or IP -
種交差性
交差種: Mouse, Rat, Human -
免疫原
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ポジティブ・コントロール
- WB: Wild type HAP1 whole cell lysate; HepG2, Jurkat, NIH/3T3, PC-12, Ramos, C6 and SH-SY5Y cell lysates; Mouse embryo, skin and lung tissue lysates; Human skin, lung and artery tissue lysates. IHC-P: Human lung carcinoma and breast carcinoma tissues.
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% 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
EP618Y -
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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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ab40759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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WB | (3) |
1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa).
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IHC-P | (4) |
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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特記事項 |
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WB
1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ターゲット情報
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機能
Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling. Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site; required for syngernistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor. -
関連疾患
Defects in SMAD4 are a cause of pancreatic cancer (PNCA) [MIM:260350].
Defects in SMAD4 are a cause of juvenile polyposis syndrome (JPS) [MIM:174900]; also known as juvenile intestinal polyposis (JIP). JPS is an autosomal dominant gastrointestinal hamartomatous polyposis syndrome in which patients are at risk for developing gastrointestinal cancers. The lesions are typified by a smooth histological appearance, predominant stroma, cystic spaces and lack of a smooth muscle core. Multiple juvenile polyps usually occur in a number of Mendelian disorders. Sometimes, these polyps occur without associated features as in JPS; here, polyps tend to occur in the large bowel and are associated with an increased risk of colon and other gastrointestinal cancers.
Defects in SMAD4 are a cause of juvenile polyposis/hereditary hemorrhagic telangiectasia syndrome (JP/HHT) [MIM:175050]. JP/HHT syndrome phenotype consists of the coexistence of juvenile polyposis (JIP) and hereditary hemorrhagic telangiectasia (HHT) [MIM:187300] in a single individual. JIP and HHT are autosomal dominant disorders with distinct and non-overlapping clinical features. The former, an inherited gastrointestinal malignancy predisposition, is caused by mutations in SMAD4 or BMPR1A, and the latter is a vascular malformation disorder caused by mutations in ENG or ACVRL1. All four genes encode proteins involved in the transforming-growth-factor-signaling pathway. Although there are reports of patients and families with phenotypes of both disorders combined, the genetic etiology of this association is unknown.
Defects in SMAD4 may be a cause of colorectal cancer (CRC) [MIM:114500]. -
配列類似性
Belongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain. -
ドメイン
The MH1 domain is required for DNA binding.
The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import. -
翻訳後修飾
Monoubiquitinated on Lys-519 by E3 ubiquitin-protein ligase TRIM33. Monoubiquitination hampers its ability to form a stable complex with activated SMAD2/3 resulting in inhibition of TGF-beta/BMP signaling cascade. Deubiqitination by USP9X restores its competence to mediate TGF-beta signaling. -
細胞内局在
Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with R-SMAD. - Information by UniProt
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参照データベース
- Entrez Gene: 4089 Human
- Entrez Gene: 17128 Mouse
- Entrez Gene: 50554 Rat
- Omim: 600993 Human
- SwissProt: Q13485 Human
- SwissProt: P97471 Mouse
- SwissProt: O70437 Rat
- Unigene: 75862 Human
see all -
別名
- (Small) Mothers Against Decapentaplegic antibody
- Deleted in Pancreatic Carcinoma 4 antibody
- Deleted in Pancreatic Carcinoma antibody
see all
画像
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: SMAD4 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 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.
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All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (purified)
Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate
Lane 2 : Ramos (Human Burkitt's lymphoma cell line) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 65 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution
Lane 1 : NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate
Lane 2 : Mouse embryo tissue lysate
Lane 3 : Mouse skin tissue lysate
Lane 4 : Mouse lung tissue lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate
Lane 6 : C6 cell lysate
Lane 7 : Rat skin tissue lysate
Lane 8 : Rat lung tissue lysate
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: 65 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution
Lane 1 : SW480 (Human colorectal adenocarcinoma cell line) cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
Lane 4 : Human skin tissue lysate
Lane 5 : Human lung tissue lysate
Lane 6 : Human artery tissue lysate
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: 65 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Different batches of ab40759 were tested on Ramos (Human Burkitt's lymphoma B lymphocyte) lysate at 0.7 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 60 kDa.
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Anti-Smad4 antibody [EP618Y] (ab40759) at 1/10000 dilution (purified) + NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 65 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
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Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (purified) + PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 65 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
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Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (unpurified) + SHSY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg
Predicted band size: 65 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
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 (153)
ab40759 は 153 報の論文で使用されています。
- Li Y et al. Long non‑coding RNA MBI‑52 inhibits the development of liver fibrosis by regulating the microRNA‑466g/SMAD4 signaling pathway. Mol Med Rep 25:N/A (2022). PubMed: 34850963
- Xue Y et al. Myokine Irisin promotes osteogenesis by activating BMP/SMAD signaling via αV integrin and regulates bone mass in mice. Int J Biol Sci 18:572-584 (2022). PubMed: 35002510
- Sun C et al. Exosomal microRNA-618 derived from mesenchymal stem cells attenuate the progression of hepatic fibrosis by targeting Smad4. Bioengineered 13:5915-5927 (2022). PubMed: 35199612
- Sherman EJ et al. Identification of cell type specific ACE2 modifiers by CRISPR screening. PLoS Pathog 18:e1010377 (2022). PubMed: 35231079
- Wu J et al. Rubus chingii Hu. unripe fruits extract ameliorates carbon tetrachloride-induced liver fibrosis and improves the associated gut microbiota imbalance. Chin Med 17:56 (2022). PubMed: 35549741