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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, ICC, IP | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE992 |
| Description | Anti-Septin-9 Antibody, clone 10C10 |
| Alternate Names |
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| Background Information | Septin-9 (UniProt Q9UHD8; also known as MLL septin-like fusion protein, MLL septin-like fusion protein MSF-A, Ovarian/Breast septin, Ov/Br septin, SeptD1, Septin D1) is encoded by the SEPT9 (also known as AF17q25, KIAA0991, MSF, MSF1, PNUTL4, SINT1) gene (Gene ID 10801) in human. Septins are GTPases capable of forming higher-order oligomers and filamentous polymers that contact with cell membrane and cytoskeleton. Septins are involved in various cellular processes, including cytokinesis, karyokinesis, exocytosis, and cell shape maintenance. Septins exist in complexes that are composed of ordered non-polar arrays of multiple different septins. Human genome contains 13 septin genes that are grouped into the SEPT2 subgroup (septins 1, 2, 4, and 5), the SEPT6 subgroup (septins 6, 8, 10, 11, and 14), the SEPT3 subgroup (septins 3, 9, and 12), and the SEPT7 subgroup (septin 7). Among mammalian septins, septin-9 undergoes the most alternative splicings, alteration in septin-9 isoforms expression has been linked to cancer and SEPT9 gene mutations have been linked to hereditary neuralgic amyotrophy. Cdk1-catalyzed septin-9 Thr24 phosphorylation is reported to promote septin-9 interaction with the peptidylprolyl isomerase Pin1. Genetic knockdown of either protein results in abscission defect during cell division. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-Septin-9 Antibody, clone 10C10 is validated for use in Western Blotting, Immunocytochemistry, Immunoprecipitation for the detection of Septin-9. |
| Key Applications |
|
| Application Notes | Immunocytochemistry Analysis: A representative lot detected septin-9 immunoreactivity in U2OS cells (Courtesy of Dr. William S. Trimble, University of Toronto, Canada). Immunocytochemistry Analysis: Clone 10C10 hybridoma culture supernatant immunostained intracellular septin-9 in hTERT-immortalized retinal pigment epithelial (hTERT-RPE) cells (Courtesy of Dr. William S. Trimble, University of Toronto, Canada). Immunocytochemistry Analysis: A representative lot detected septin-9 immunoreactivity colocalized with septin-2 and septin-11 at the cleavage furrow in anaphase and the midbody in telophase HeLa cells (Estey, M.P., et al. (2010). J. Cell Biol. 191(4):741-749). Immunoprecipitation Analysis: A representative lot immunoprecipited septin-9 from mitotically arrested HeLa cells following nocodazole (Cat. No. 487928) treatment. Cdk1 inhibition by Roscovitine (Cat. No. 57360) treatment or siRNA-meidated Cdk1 knowndown diminished the phosphroylation of the immunoprecipitated septin-9 (Estey, M.P., et al. (2013). J. Biol. Chem. 288(42):30075-30086). Immunoprecipitation Analysis: A representative lot immunoprecipitated significantly more septin-9 from unsynchronized than from mitotically synchronized HeLa cells, the difference was normalized following siRNA-mediated Pin1 knockdown (Estey, M.P., et al. (2013). J. Biol. Chem. 288(42):30075-30086). Western Blotting Analysis: A representative lot specifically detected recombinant septin-9 spliced isoforms and fragments that contain the epitope region sequence, but not recombinant isoforms or fragments lacking the epitope (Estey, M.P., et al. (2013). J. Biol. Chem. 288(42):30075-30086). Western Blotting Analysis: A representative lot specifically detected purified recombinant human septin-9, but not purified recombinant spetin-1, -2, -5, -6, -7, -8. -9. -11, -12 (Estey, M.P., et al. (2010). J. Cell Biol. 191(4):741-749). Western Blotting Analysis: A representative lot detected the presence of co-precipitated septin-9 in septin-2 immunoprecipitates from HeLa cell lysates (Estey, M.P., et al. (2010). J. Cell Biol. 191(4):741-749). Note: For Western blotting application, prior enrichment of septin-9 by immunoprecipitation or other purification methods is highly recommended. Clone 10C10 is not recommended for probing septin-9 using whole cell lysates. |
| Biological Information | |
|---|---|
| Immunogen | His-tagged recombinant human septin-9. |
| Epitope | Internal (N-terminal half). |
| Clone | 10C10 |
| Concentration | Please refer to lot specific datasheet. |
| Host | Mouse |
| Specificity | Clone 10C10 specifically detected purified recombinant human Septin-9, but not recombinant Spetin-1, -2, -5, -6, -7, -8. -9. -11, or -12, by Western blotting (Estey, M.P., et al. (2010). J. Cell Biol. 191(4):741-749). The epitope has been mapped to region present in human Septin-9 spliced isoforms 1 (Epsilon, MSF-A), 2 (Alpha), 5 (Gamma), and 7, but absent in spliced isoforms 3 (Beta, MSF-B), 4 (Delta), 8, and 9 reported by UniProt (Q9UHD8). |
| Isotype | IgG1κ |
| Species Reactivity |
|
| 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 | ~69 kDa observed. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting of recombinant human septin-9. Western Blotting Analysis: 0.5 µg/mL of this antibody detected His-tagged recombinant human septin-9. |
| 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 μg |