Human Smad4 Antibody

Catalog # Availability Size / Price Qty
AF2097
AF2097-SP
Detection of Human Smad4 by Western Blot.
3 Images
Product Details
Citations (4)
FAQs
Supplemental Products
Reviews (1)

Human Smad4 Antibody Summary

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

Western Blot Detection of Human Smad4 by Western Blot. View Larger

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. View Larger

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. View Larger

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

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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.

4 Citations: Showing 1 - 4
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  1. 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
  2. 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
  3. 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
  4. 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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Human Smad4 Antibody
By Anonymous on 03/16/2017
Application: WB Sample Tested: Breast cancer cells Species: Human

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.