| 货号 | AF2097-SP |
| 别名 | 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 | 全称 | Mothers Against DPP Homolog 4 |
| 反应种属 | Human |
| 应用 | Western Blot(0.25 µg/mL) Chromatin Immunoprecipitation (ChIP)(5 µg/5 x 106cells) Immunocytochemistry(5-15 µg/mL) |
| 目标/特异性 | 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. |
| 使用方法 | Western Blot: 0.25 µg/mL Chromatin Immunoprecipitation (ChIP): 5 µg/5 x 106cells Immunocytochemistry: 5-15 µg/mL |
| 来源 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 产品组分 |
| 供应商 | R&D Systems |
| Entrez Gene IDs | 4089 (Human) |
| 应用文献 | |
| 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. Chromatin and transcriptional signatures for Nodal signaling during endoderm formation in hESCs. | |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | E. coli-derived recombinant human Smad4 Pro139-Asp332 Accession # Q13485 |
| 生物活性 | Human |
| 标记 | Unconjugated |
| 溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 背景 | Smads are a family of intracellular proteins that transmit transforminggrowth factor beta (TGF-beta ) superfamily signals from the cell surface tothe nucleus. Upon signal-induced phosphorylation, Smad subunitsassociate with the common-mediator subunit, Smad4. This heteromericcomplex then translocates into the nucleus to exert transcriptionalcomodulator activity. |
| 运输条件 | Blue Ice |
| 存放说明 | 4℃ |
| 参考文献 |
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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. | |
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. | |
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. |
