Name: Anti-Smad4 antibody [EP618Y]
Brand: RabMAb
SKU: ab40759
Rating: 4 (9 reviews)

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Western blot - Anti-Smad4 antibody [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

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Carrier Free HRP

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

Synthetic peptide within Human Smad4 aa 500 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: Q13485
(Peptide available as ab228416)
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.
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. 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...
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

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Smad4 protein (ab81764)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10 - 245 kDa) (ab116028)

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
WB	(3)	1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa).
IHC-P	(4)	1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.

特記事項
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. See IHC antigen retrieval protocols.

追加情報

Is unsuitable for Flow Cyt,ICC/IF or IP.

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.
Target information above from: UniProt accession Q13485 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: 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
- Unigene: 100399 Mouse
- Unigene: 9774 Rat
see all
別名
- (Small) Mothers Against Decapentaplegic antibody
- Deleted in Pancreatic Carcinoma 4 antibody
- Deleted in Pancreatic Carcinoma antibody
- Deleted in pancreatic carcinoma locus 4 antibody
- Deletion target in pancreatic carcinoma 4 antibody
- DPC 4 antibody
- DPC4 antibody
- hSMAD4 antibody
- JIP antibody
- MAD homolog 4 antibody
- MAD mothers against decapentaplegic Drosophila homolog 4 antibody
- MAD mothers against decapentaplegic homolog 4 antibody
- MADH 4 antibody
- MADH4 antibody
- Med antibody
- Medea antibody
- Mothers against decapentaplegic homolog 4 antibody
- Mothers against decapentaplegic, Drosophila, homolog of, 4 antibody
- Mothers against DPP homolog 4 antibody
- MYHRS antibody
- OTTHUMP00000163548 antibody
- SMA- and MAD-related protein 4 antibody
- SMAD 4 antibody
- SMAD family member 4 antibody
- SMAD mothers against DPP homolog 4 antibody
- SMAD4 antibody
- SMAD4_HUMAN antibody
see all

Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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 minutes

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

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) - Anti-Smad4 antibody [EP618Y] (ab40759)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)This image is courtesy of an anonymous Abreview

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.
See Abreview
Anti-Smad4 antibody [EP618Y] (ab40759)

Abreview protocol for ChIP using ab40759
Recommended FACS protocol

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab40759 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。

ab40759 は 170 報の論文で使用されています。

Luo D et al. Pirfenidone suppressed triple-negative breast cancer metastasis by inhibiting the activity of the TGF-β/SMAD pathway. J Cell Mol Med 27:456-469 (2023). PubMed: 36651490
Li W et al. circRNA ITGA7 restrains growth and enhances radiosensitivity by up-regulating SMAD4 in colorectal carcinoma. Open Med (Wars) 18:20220604 (2023). PubMed: 36694626
Selven H et al. High Expression of IRS-1, RUNX3 and SMAD4 Are Positive Prognostic Factors in Stage I-III Colon Cancer. Cancers (Basel) 15:N/A (2023). PubMed: 36900240
Ma Z et al. Somatic-to-primordial germ cell-like transformation is critical in tumor initiation of mouse breast tumor 4T1 cells. Am J Cancer Res 13:1471-1485 (2023). PubMed: 37168326
Wu W et al. Liver X receptors and estrogen receptor β, two players in a rare subtype of NSCLC. Int J Biol Sci 19:2848-2859 (2023). PubMed: 37324952

View all Publications for this product

レビューと Q&A

Show All レビュー Q&A

レビューを送る質問を送る

1-9 of 9 Abreviews or Q&A

Application

Western blot

Sample

Coturnix japonica Tissue lysate - whole (yolk sac membrane from embryonic day 3 embryos)

Gel Running Conditions

Non-reduced Non-Denaturing (Native) (10% gel)

Loading amount

50 µg

Specification

yolk sac membrane from embryonic day 3 embryos

Blocking step

Gelatin buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · 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.

MISS. HAN-JEN LIN

Verified customer

投稿 Apr 17 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Liver)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Dako

Permeabilization

Specification

Liver

Fixative

Formaldehyde

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.

Abcam user community

Verified customer

投稿 Jun 01 2016

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (postnatal day 21 mouse ovary)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 0.01M citric acide/sodium citrate

Permeabilization

Specification

postnatal day 21 mouse ovary

Blocking step

BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

4% PFA

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.

Abcam user community

Verified customer

投稿 Jan 05 2015

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (A549)

Permeabilization

Yes - 0.25% Triton X-100/PBS

Specification

A549

Fixative

Paraformaldehyde

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.

Abcam user community

Verified customer

投稿 Apr 11 2013

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Lung tissue)

Specification

Lung tissue

Fixative

HOPE

Antigen retrieval step

None

Permeabilization

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.

Abcam user community

Verified customer

投稿 Mar 27 2013

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rat Tissue sections (femur)

Specification

femur

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrated buffer, pH6.0

Permeabilization

Blocking step

BSA as blocking agent for 20 minute(s) · Concentration: 1% · 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.

Abcam user community

Verified customer

投稿 Aug 23 2012

Application

Western blot

Sample

Human Cell lysate - whole cell (lymphoblats)

Loading amount

40 µg

Specification

lymphoblats

Gel Running Conditions

Reduced Denaturing (4-12% Bis-Tris Gel, MOPS Buffer)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°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.

DR. Lukas Rambousek

Verified customer

投稿 Dec 15 2010

Application

ChIP

Sample

Human Cell lysate - whole cell (MDA-MB-231 Cells treated with 2 ng/µl TGF-beta1)

Negative control

Polyclonal anti-HA antibody (for detection of non-specific binding). The graph is shown as a "fold enrichment" relative to the average detected HA binding.

Specification

MDA-MB-231 Cells treated with 2 ng/µl TGF-beta1

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde

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.

Abcam user community

Verified customer

投稿 Feb 14 2007

Application

Western blot

Sample

Human Cell lysate - whole cell (colon cancer)

Loading amount

50 µg

Specification

colon cancer

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

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.

Abcam user community

Verified customer

投稿 Dec 11 2006

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

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

Isotype control

Recombinant Protein

Related Products

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

関連疾患

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

参考文献 (170)

レビューと Q&A

Western blot abreview for Anti-Smad4 antibody [EP618Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Smad4 antibody [EP618Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad4 antibody [EP618Y]

Western blot abreview for Anti-Smad4 antibody [EP618Y]

ChIP abreview for Anti-Smad4 antibody [EP618Y]

Western blot abreview for Anti-Smad4 antibody [EP618Y]