Human Smad4 Antibody

(3 citations)
(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human Smad4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 15% cross-reactivity with recombinant human (rh) Smad1 and less than 5% cross-reactivity with rhSmad9, rhSmad5, and rhSmad7 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Smad4
    Pro139-Asp332
    Accession # Q13485
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25 µg/mL
    See below
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/5 x 106 cells
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
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.
Data Examples
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.
Chromatin Immunoprecipitation (ChIP)
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. The p21 promoter was detected by standard PCR.
Immunocytochemistry
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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.
  • Long Name:
    Mothers Against DPP Homolog 4
  • Entrez Gene IDs:
    4089 (Human)
  • Alternate Names:
    DPC4; DPC4SMAD, mothers against DPP homolog 4 (Drosophila); hSMAD4; JIP; MAD homolog 4; MADH4; MADH4mothers against decapentaplegic homolog 4; mothers against decapentaplegic homolog 4; mothers against decapentaplegic, Drosophila, homolog of, 4; SMAD 4; SMAD family member 4MAD, mothers against decapentaplegic homolog 4 (Drosophila); SMAD, mothers against DPP homolog 4; Smad4
Related Research Areas
Citations:

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.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. 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 Type: Saliva
    Application: WB
  2. 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 Type: Cell Lysates
    Application: WB
  3. 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 Type: Cell Lysates
    Application: ChIP
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Goat HRP-DAB Cell & Tissue Staining Kit

CTS008 13
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Isotype Controls
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Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
Description Application Cat# Citations Images  

Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Western Blot Human Smad4 Antibody AF2097
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Excellent
 WB Human Anonymous 03/16/2017
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Western Blot Human Smad4 Antibody AF2097
Western Blot: Human Smad4 Antibody [AF2097]

Summary

ApplicationWestern Blot
Sample TestedBreast cancer cells
SpeciesHuman

Other Experimental Details

Other Experimental DetailsWestern 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.

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