SMAD4 antibody
Principal name
SMAD4 antibody
Alternative names for SMAD4 antibody
SMAD family member 4, SMAD-4, SMAD 4, MADH4, MAD homolog 4, DPC4, Mothers against decapentaplegic homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4
Ncbi ID
NP_005350.1, 9606, NP_005350, 10090, ABM82766.1, NM_008540, NM_019275, NM_005359
Available reactivities
Available hosts
Available applications
Western blot / Immunoblot (WB), Enzyme Immunoassay (E), Immunocytochemistry/Immunofluorescence (ICC/IF), Paraffin Sections (P), Dot blot (Dot), Immunoprecipitation (IP)
Background of SMAD4 antibody
SMAD4 (also known as Mothers against decapentaplegic homolog 4, Mothers against DPP homolog 4, deletion target in pancreatic carcinoma 4 and hSMAD4) is a common mediator of signal transduction by TGF-beta (transforming growth factor), but is also involved in cancer development and metastases as a tumor suppressor. SMAD4 promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. SMAD4 may form trimers with receptor-regulated SMAD (R-SMAD) and interacts with ATF2, COPS5, DACH1, MSG1, SKI and TRIM33. In the absence of ligand SMAD4 is found in the cytoplasm, but when complexed with R-SMAD, translocates to the nucleus. Defects in SMAD4 are a cause of pancreatic carcinoma and juvenile polyposis syndrome (JPS), a syndrome in which patients are at risk for developing gastrointestinal cancers.
General readings
1. Kim, B.-G.; Li, C.; Qiao, W.; Mamura, M.; Kasprzak, B.; Anver, M.; Wolfraim, L.; Hong, S.; Mushinski, E.; Potter, M.; Kim, S.-J.; Fu, X.-Y.; Deng, C.; Letterio, J. J. : Smad4 signalling in T cells is required for suppression of gastrointestinal cancer. Nature 441: 1015-1019, 2006. Note: Erratum: Nature 444: 780 only, 2006.
2. Houlston, R.; Bevan, S.; Williams, A.; Young, J.; Dunlop, M.; Rozen, P.; Eng, C.; Markie, D.; Woodford-Richens, K.; Rodriguez-Bigas, M. A.; Leggett, B.; Neale, K.; Phillips, R.; Sheridan, E.; Hodgson, S.; Iwama, T.; Eccles, D.; Bodmer, W.; Tomlinson, I. : Mutations in DPC4 (SMAD4) cause juvenile polyposis syndrome, but only account for a minority of cases. Hum. Molec. Genet. 7: 1907-1912, 1998.
3 Howe, J. R.; Roth, S.; Ringold, J. C.; Summers, R. W.; Jarvinen, H. J.; Sistonen, P.; Tomlinson, I. P. M.; Houlston, R. S.; Bevan, S.; Mitros, F. A.; Stone, E. M.; Aaltonen, L. A. : Mutations in the SMAD4/DPC4 gene in juvenile polyposis. Science 280: 1086-1088, 1998.
Immunofluorescence of monoclonal antibody to SMAD4 on HeLa cell. [antibody concentration 10 ug/ml]
SMAD4 monoclonal antibody (M01), clone 3B9 Western Blot analysis of SMAD4 expression in Hela NE ( Cat # L013V3 ).
Proximity Ligation Analysis (PLA) of protein-protein interactions between CDKN1A and SMAD4 HeLa cells were stained with anti-CDKN1A rabbit purified polyclonal 1:1200 and anti-SMAD4 mouse monoclonal antibody 1:50. Signals were detected by Duolink® 30 Detection Kit 613(red), and nuclei were counterstained with DAPI (blue). Each red dot represents the detection of protein-protein interaction complex.
Immunofluorescence of monoclonal antibody to SMAD4 on HeLa cell. [antibody concentration 10 ug/ml]
SMAD4 monoclonal antibody (M02), clone 3E2. Western Blot analysis of SMAD4 expression in K-562 ( Cat # L009V1 ).
SMAD4 monoclonal antibody (M02), clone 3E2. Western Blot analysis of SMAD4 expression in IMR-32 ( Cat # L008V1 ).
SMAD4 monoclonal antibody (M02), clone 3E2 Western Blot analysis of SMAD4 expression in Hela NE ( Cat # L013V3 ).
Detection limit for recombinant GST tagged SMAD4 is approximately 3ng/ml as a capture antibody.
Proximity Ligation Analysis (PLA) of protein-protein interactions between AKT1 and SMAD4. Huh7 cells were stained with anti-AKT1 rabbit purified polyclonal 1:1200 and anti-SMAD4 mouse monoclonal antibody 1:50. Signals were detected by Duolink® 30 Detection Kit 613(red), and nuclei were counterstained with DAPI (blue). Each red dot represents the detection of protein-protein interaction complex.
Detection limit for recombinant GST tagged SMAD4 is approximately 3ng/ml as a capture antibody.
SMAD4 polyclonal antibody (A01), Lot # 051017JC01 Western Blot analysis of SMAD4 expression in K-562 ( Cat # L009V1 ).
Human U2OS; . Sample Type: Human nuclear cell extracts (30ug). Primary Dilution: 1:1000. Secondary Antibody: anti-Rabbit HRP. Secondary Dilution: 1:20000. Image Submitted By: Katarina Luciakova. Cancer Research Institute . See Customer Feedback tab for detailed information.. .; SMAD4 antibody - middle region (ARP32005_T100) in Human U2OS cells using Western Blot
Human U2OS; SMAD4 antibody - middle region (ARP32005_T100) validated by WB using U2OS nuclear extracts; SMAD4 antibody - middle region (ARP32005_T100) in Human U2OS cells using Western Blot
Human HepG2; WB Suggested Anti-SMAD4 Antibody Titration: 1.25ug/ml. ELISA Titer: 1:312500. Positive Control: HepG2 cell lysate; SMAD4 antibody - middle region (ARP32005_T100) in Human HepG2 cells using Western Blot
Immunohistochemistry (IHC) analyzes of Smad4 antibody (Cat.-No.: AP06725PU-N) in paraffin-embedded human brast carcinoma tissue.
Western blot (WB) analysis of Smad4 antibody (Cat.-No.: AP06725PU-N) in extracts from HeLa cells.
Western blot (WB) analysis of Smad4 antibody (Cat.-No.: AP06725PU-N) at 1/500 dilution:
Lane 1: A549 whole cell lysate.
Lane 2: Raw264.7 whole cell lysate.
Lane 3: PC12 whole cell lysate.
Lane 1: A549 whole cell lysate.
Lane 2: Raw264.7 whole cell lysate.
Lane 3: PC12 whole cell lysate.
Formalin-fixed and paraffin-embedded human breast cancer tissue reacted with SMAD4 (phospho T277) polyclonal antibody ( Cat # PAB1965 ) which was peroxidase-conjugated to the secondary antibody followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Dot blot analysis of SMAD4 (phospho T277) polyclonal antibody ( Cat # PAB1965 ) on nitrocellulose membrane. 50 µg of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5 µg/ml.
SMAD4 polyclonal antibody ( Cat # PAB6049 ) staining ( 0.5 µg/ml ) of NIH/3T3 cell lysate ( RIPA buffer, 35 µg total protein per lane ) . Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
SMAD4 monoclonal antibody (M01), clone 3B9 Western Blot analysis of SMAD4 expression in Hela NE ( Cat # L013V3 ).
Recombinant human SMAD4, aa 1-552 , His-tagged: 15% SDS-PAGE (3ug)
Formalin-fixed and paraffin-embedded Breast Carcinoma reacted with SMAD4 pThr277 (center) (Cat.#AP12711PU-N), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
Dot blot analysis of anti-Phospho-SMAD4 pThr277 Antibody (Cat#AP12711PU-N) on nitrocellulose membrane. 50ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5ug per ml.
Western blot using SMAD4 polyclonal antibody ( Cat # PAB11265 ) to detect over-expressed SMAD4 in transfected COS cells ( lane 2 ).Lane 1 contains lysate from mock transfected COS cells.A doublet band is seen in the SMAD4 transfected lysate with the upper band ( arrowhead ) representing SMAD4 and the lower band being non-specific staining.The primary antibody at a 1:1,000 dilution.Personal Communication, Kathleen Flanders, CCR-NCI, Bethesda, MD.
Western blot using affinity purified anti-SMAD4 to detect over-expressed SMAD4 in transfected COS cells (lane 2). Lane 1 contains lysate from mock transfected COS cells. A doublet band is seen in the SMAD4 transfected lysate with the upper band (arrowhead) representing SMAD4 and the lower band being non-specific staining. The membrane was probed with tprimary antibody at a 1:1,000 dilution.
Human Lung; . Rabbit Anti-SMAD4 antibody. . Catalog Number:. ARP38605_P050 . Paraffin Embedded Tissue: Human Lung cell . Cellular Data: Epithelial cells of renal tubule. Antibody Concentration: 4.0-8.0 ug/ml. Magnification: 400X; SMAD4 antibody (ARP38605_P050) in Human Lung cells using Immunohistochemistry
Human 293T; WB Suggested Anti-SMAD4 Antibody Titration: 0.2-1 ug/ml. ELISA Titer: 1:312500. Positive Control: 293T cell lysate. SMAD4 is supported by BioGPS gene expression data to be expressed in HEK293T.; SMAD4 antibody - N-terminal region (ARP38605_P050) in Human 293T cells using Western Blot
Mouse Brain; WB Suggested Anti-Smad4 Antibody Titration: 0.2-1 ug/ml. ELISA Titer: 1:312500. Positive Control: Mouse Brain; Smad4 antibody - middle region (ARP32688_P050) in Mouse Brain cells using Western Blot
Immunohistochemical analysis of paraffin-embedded lung cancer tissues using SMAD4 antibody Cat.-No AM06501SU-N with DAB staining.
Immunofluorescence analysis of NIH/3T3 cells using SMAD4 antibody Cat.-No AM06501SU-N (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.
Western blot analysis using SMAD4 antibody Cat.-No AM06501SU-N against A431 (1), SK-N-SH (2), K562 (3), HepG2 (4) and HUVE12 (5) cell lysate.
Flow Cytometric analysis of K562 cells using SMAD4 antibody Cat.-No AM06501SU-N (green) and negative control (purple).
Immunohistochemical analysis of SMAD4 staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody 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.
Immunofluorescent analysis of SMAD4 staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).
Western blot analysis of SMAD4 expression in HepG2 (A); HeLa (B); NIH3T3 (C); COS7 (D) whole cell lysates.
Western blot analysis of SMAD4 Antibody (T277)(arrow) using rabbit polyclonal SMAD4 Antibody (T277) (Cat.#TA302055). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the SMAD4 gene (Lane 2) (Origene Technologies).
Confocal immunofluorescent analysis of SMAD4 Antibody (T277)(Cat#TA302055) with Hela cell followed by Alexa Fluor 488-conjugated goat anti-rabbit lgG (green). Actin filaments have been labeled with Alexa Fluor 555 phalloidin (red).DAPI was used to stain the cell nuclear (blue).
TA302724 staining (0.5µg/ml) of NIH-3T3 cell lysate (RIPA buffer, 35µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
Predicted cell location: Cytoplasm. Positive control: Human ovarian cancer tissue. Recommended dilution: 1/25-100 The image on the left is immunohistochemistry of paraffin-embedded Human ovarian cancer tissue using SMAD4 antibody at dilution 1/30, on the right is treated with the fusion protein. (Original magnification: x200)
Western blot analysis of SMAD4 (arrow) using rabbit polyclonal SMAD4 Antibody (C-term) (Cat. #TA324984). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the SMAD4 gene (Lane 2) .
Formalin-fixed and paraffin-embedded human skeletal muscle reacted with SMAD4 Antibody (C-term), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
IF image of Hela cell stained with SMAD4 Antibody (C-term)(Cat#TA324984).Hela cells were incubated with SMAD4 primary antibody (1:25, 1 h at 37?). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:400).Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7 units/ml). Nuclei were counterstained with DAPI (blue) . SMAD4 immunoreactivity is localized to Cytoplasm and Nucleus significantly.
SMAD4 Antibody (C-term) (Cat. #TA324984) flow cytometric analysis of MCF-7 cells (bottom histogram) compared to a negative control cell (top histogram).FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.
Western blot analysis of extracts from HeLa cells, COS-7 cells, HepG2 cells and HUVEC cells, using Smad4 antibody.The lane on the right is treated with the synthesized peptide.
WB Suggested Anti-SMAD4 Antibody Titration: 1.25 ug/ml; ELISA Titer: 1:312500; Positive Control: HepG2 cell lysate
Sample Type: Human nuclear cell extracts (30 ug); Primary Dilution: 1:1000; Secondary Antibody: anti-Rabbit HRP; Secondary Dilution: 1:20000; Image Submitted By: Katarina Luciakova; Cancer Research Institute See Customer Feedback tab for detailed informat
SMAD4 antibody - middle region validated by WB using U2OS nuclear extracts
WB Suggested Anti-Smad4 Antibody Titration: 0.2-1 ug/ml; ELISA Titer: 1:312500; Positive Control: Mouse Brain
Western blot analysis of extracts of various cell lines, using SMAD4 antibody.
Immunofluorescence analysis of U20S cell using SMAD4 antibody. Blue: DAPI for nuclear staining.
WB Suggested Anti-SMAD4 Antibody Titration: 0.2-1 ug/ml; ELISA Titer: 1: 312500; Positive Control: 293T cell lysate; SMAD4 is supported by BioGPS gene expression data to be expressed in HEK293T
Rabbit Anti-SMAD4 antibody; Paraffin Embedded Tissue: Human Lung cell; Cellular Data: Epithelial cells of renal tubule; Antibody Concentration: 4.0-8.0 ug/ml; Magnification: 400X
Host: Rabbit; Target Name: Smad4; Sample Tissue: Mouse Testis lysates; Antibody Dilution: 1.0 ug/ml
Western blot analysis on HeLa cell lysate using Smad4 Antibody
Primary Antibodies
| Catalog No. | Host | Iso. | Clone | Pres. | React. | Applications | |
|---|---|---|---|---|---|---|---|
| TA302724 | SMAD4 (C-term) antibody |
||||||
| Goat Polyclonal Antibody against SMAD4 | |||||||
 |
Goat | Aff - Purified | Bov, Can, Hu, Ms, Por, Rt | E, WB |
0.1 mg /
$360.00
|
||
| OriGene Technologies, Inc. | |||||||