Human Smad4 Antibody Summary
Pro139-Asp332
Accession # Q13485
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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

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.

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.

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 - 5
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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;33(2):108253.
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;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;40(8):1183-1191.
Species: Human
Sample Types: Saliva
Applications: 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;7(4):e1082.
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation -
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;357(2):492-504.
Species: Human
Sample Types: Cell Lysates
Applications: ChIP
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