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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | WB, IH(P), ChIP, ICC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE1057 |
| Description | Anti-ADA3 Antibody, clone 5C9/C8 |
| Alternate Names |
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| Background Information | Transcriptional adapter 3 (UniProt O75528; also known as ADA3 homolog, ADA3-like protein, Alteration/deficiency in activation 3, hADA3, STAF54, Transcriptional adapter 3-like) is encoded by the TADA3 (also known as ADA3, TADA3L) gene (Gene ID 10474) in human. Alteration/deficiency in activation 3 (ADA3) is a component of several transcriptional co-activator and histone acetyltransferase (HAT) complexes and plays a critical role in in cell cycle regulation. Ada3 deletion in mice is embryonic lethal, and Ada3-deficient mouse embryonic fibroblasts (MEFs) exhiit a severe proliferation defect, dramatic changes in global histone acetylation, mitotic defects, as well as a delay in G2/M-to-G1 and G1-to-S transition. ADA3 also plays a role in genomic stability by controlling DNA repair checkpoints. ChIP-seq analsis reveals that ADA3 is significantly associated with human centromere regions across most chromosomes. In addition, ADA3 is found associated with CENP-B throughout all phases of the cell cycle, and CENP-B centromere binding decreased upon ADA3 knockdown. Wild-type human ADA3, but not CENP-B-binding deficient ADA3 mutant, prevented cell proliferation defects in MEFs following endogenous mouse ADA3 knockown. ADA3 overexpression and mislocalization correlates with poor prognosis in breast cancer patients. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-ADA3, clone 5C9/C8, Cat. No. MABE1057, is a highly specific mouse monoclonal antibody that targets TADA3 and has been tested in Chromatin Immunoprecipitation (ChIP), Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Key Applications |
|
| Application Notes | Immunohistochemistry Analysis: A representative lot detected nuclear ADA3 immunoreactivity in formalin-fixed, paraffin-embedded human breast carcinoma tissue section (Courtesy of Vimla Band, Ph.D., University of Nebraska USA). Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected ADA3 occupancy at the X chromosome centromere HOR region, the kinetochore assembly site also occupied by CENP-A and CENP-B, using TERT-immortalized human mammary epithelial cell 76N-TERT chromatin preparation. shRNA-mediated ADA3 knockdown led to a reduction in CENP-B, but not CENP-A, recruitment to the HOR region (Mohibi, S., et al. (2015). J. Biol. Chem. 290(47):28299-28310). Immunocytochemistry Analysis: A representative lot detected ADA3 nuclear immunoreactivity in untransfected, but not ADA3 shRNA-transfected, TERT-immortalized human mammary epithelial 76N-TERT cells by fluorescent immunocytochemistry staining of 4% formaldehyde-fixed cells. Dual fluorescent staining revealed ADA3 and CENP-B nuclear colocalization (Mohibi, S., et al. (2015). J. Biol. Chem. 290(47):28299-28310). Western Blotting Analysis: Representative lots detected both the endogenous mouse ADA3 (mADA3) and the exogenously expressed human ADA3 (hADA3) in Ada3FL/FL MEFs virally infected to express FLAG-tagged full-length or a.a. 111-432 hADA3 constructs. Cre recombinase expression downregulated only mADA3, but not hADA3 (Mohibi, S., et al. (2015). J. Biol. Chem. 290(47):28299-28310; Mohibi, S., et al. (2012). J. Biol. Chem. 287(35):29442-29456). Western Blotting Analysis: A representative lot detected ADA3 expression levels among a panel of mouse tissues (Mohibi, S., et al. (2012). J. Biol. Chem. 287(35):29442-29456). |
| Biological Information | |
|---|---|
| Immunogen | Full-length recombinant human ADA3. |
| Clone | 5C9/C8 |
| Concentration | Please refer to lot specific datasheet. |
| Host | Mouse |
| Specificity | Clone 5C9/C8 specifically detected Cre recombinase expression-induced ADA3 downregulation in Ada3FL/FL MEFs. Clone 5C9/C8 immunostained the nuclei of untransfected, but not ADA3 shRNA-transfected, 76N-TERT human mammary epithelial cells (Mohibi, S., et al. (2015). J. Biol. Chem. 290(47):28299-28310; Mohibi, S., et al. (2012). J. Biol. Chem. 287(35):29442-29456). |
| Isotype | IgG1κ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G purified. |
| UniProt Number |
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| Molecular Weight | ~55 kDa observed. 48.90 kDa (human and mouse isoform 1) calculated. The larger-than-calculated band size is consistent with that reported in the literature (Mohibi, S., et al. (2015). J. Biol. Chem. 290(47):28299-28310). Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in MCF7 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected ADA3 in 10 µg of MCF-7 cell lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |