Human Smad4 Antibody Summary
Pro139-Asp332
Accession # Q13485
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human Smad4 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Smad4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2097) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Smad4 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Detection of Smad4-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 10 ng/mL Recombinant Human IL-12 (Catalog # 219-IL) overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Smad4/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human Smad4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2097) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thep21promoter was detected by standard PCR.
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Smad4 in Human PBMCs. Smad4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human Smad4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2097) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Smad4
Smads are a family of intracellular proteins that transmit transforming growth factor beta (TGF-beta ) superfamily signals from the cell surface to the nucleus. Upon signal-induced phosphorylation, Smad subunits associate with the common-mediator subunit, Smad4. This heteromeric complex then translocates into the nucleus to exert transcriptional comodulator activity.
Product Datasheets
Citations for Human Smad4 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 6
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Bone morphogenetic protein (BMP) signaling determines neuroblastoma cell fate and sensitivity to retinoic acid
Authors: Pan, M;Zhang, Y;Wright, WC;Liu, X;Passaia, B;Currier, D;Low, J;Chapple, RH;Steele, JA;Connelly, JP;Lu, M;Lee, HM;Loughran, AJ;Yang, L;Abraham, BJ;Pruett-Miller, SM;Freeman, B;Campbell, GE;Dyer, MA;Chen, T;Stewart, E;Koo, S;Sheppard, H;Easton, J;Geeleher, P;
bioRxiv : the preprint server for biology
Species: Human
Sample Types: Chromatin
Applications: Chromatin Immunoprecipitation (ChIP) -
PAI-1-Dependent Inactivation of SMAD4-Modulated Junction and Adhesion Complex in Obese Endometrial Cancer
Authors: LL Lin, ER Kost, CL Lin, P Valente, CM Wang, MG Kolonin, AC Daquinag, X Tan, N Lucio, CN Hung, CP Wang, NB Kirma, TH Huang
Cell Rep, 2020-10-13;33(2):108253.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: ICC, Immunoprecipitation, Western Blot -
An interaction network of mental disorder proteins in neural stem cells
Authors: MJ Moen, HH Adams, JH Brandsma, DH Dekkers, U Akinci, S Karkampoun, M Quevedo, CE Kockx, Z Ozgür, WF van IJcken, J Demmers, RA Poot
Transl Psychiatry, 2017-04-04;7(4):e1082.
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation -
SMAD3 and SMAD4 have a more dominant role than SMAD2 in TGF?-induced chondrogenic differentiation of bone marrow-derived mesenchymal stem cells
Authors: LM de Kroon, R Narcisi, GG van den Ak, EL Vitters, EN Blaney Dav, GJ van Osch, PM van der Kr
Sci Rep, 2017-02-27;7(0):43164.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Characterization of Membrane Integrity and Morphological Stability of Human Salivary Exosomes
Authors: N Kumeda, Y Ogawa, Y Akimoto, H Kawakami, M Tsujimoto, R Yanoshita
Biol. Pharm. Bull., 2017-01-01;40(8):1183-1191.
Species: Human
Sample Types: Saliva
Applications: Western Blot -
Chromatin and transcriptional signatures for Nodal signaling during endoderm formation in hESCs.
Authors: Kim SW, Yoon SJ, Chuong E, Oyolu C, Wills AE, Gupta R, Baker J
Dev. Biol., 2011-06-29;357(2):492-504.
Species: Human
Sample Types: Cell Lysates
Applications: ChIP
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Western Blot of breast cancer cell lysates. The PVDF membrane was probed with 1:1,000 Goat anti-Human Smad4 antibody (#AF2097) followed by 1:5,000 anti-Goat secondary antibody.
