| 货号 | 68309T |
| 目标/特异性 | Notch1 (D1E11) XP® Rabbit mAb detects intracellular epitopes between 2400 and 2500 amino acids of human Notch1. It recognizes both the full-length (~300 kDa) and the NTM region (~120 kDa). The antibody cannot detect the extracellular (ligand-binding) domain of Notch1 following cleavage at the S2 site by ADAM-type metalloproteases. Cleaved Notch1 (V1744) (D3B8) Rabbit mAb detects endogenous levels of the Notch1 intracellular domain (NICD) only when released by cleavage between Gly1753 and Val1754 (equivalent to Gly1743/Val1744 of murine notch1). The antibody does not recognize full-length Notch1 or Notch1 cleaved at other positions. The size of the NICD varies among cell lines due to mutations in the Notch1 C-terminus. RBPSUH (D10A4) XP® Rabbit mAb recognizes endogenous levels of total RBPSUH protein. MAML1 (D3K7B) Rabbit mAb recognizes endogenous levels of total MAML1 protein. This antibody does not detect MAML2 or MAML3. c-Myc (D3N8F) Rabbit mAb recognizes endogenous levels of total c-Myc protein. p21 Waf1/Cip1 (12D1) Rabbit mAb detects endogenous levels of total p21 protein. The antibody does not cross-react with other CDK inhibitors. HES1 (D6P2U) Rabbit mAb recognizes endogenous levels of total HES1 protein. Cyclin D3 (DCS22) Mouse mAb detects endogenous levels of total cyclin D3 protein. The antibody does not cross-react with cyclin D1 or cyclin D2. |
| 供应商 | CST |
| 背景 | Notch proteins (Notch1-4) are a family of transmembrane receptors that play important roles in development and the determination of cell fate (1). Mature Notch receptors are processed and assembled as heterodimeric proteins, with each dimer comprised of a large extracellular ligand-binding domain, a single-pass transmembrane domain, and a smaller cytoplasmic subunit (Notch intracellular domain, NICD) (2). Binding of Notch receptors to ligands of the Delta-Serrate-Lag2 (DSL) family triggers heterodimer dissociation, exposing the receptors to proteolytic cleavages; these result in release of the NICD, which translocates to the nucleus and activates transcription of downstream target genes (3,4). RBPSUH (Recombining Binding Protein, SUppressor of Hairless), is the DNA-binding component of the transcription complex regulated by canonical Notch signaling. Binding of Notch with RBPSUH activates a transcription activation complex that includes Mastermind-like (MAML) proteins, leading to transcriptional activation of Notch target genes (5-7). The NICD binds and activates c-Myc which functions as a transcriptional regulator with roles in various aspects of cell behavior including proliferation, differentiation and apoptosis (8). The tumor suppressor protein p21 Waf1/Cip1 acts as an inhibitor of cell cycle progression. The NICD-RBPSUH complex binds and activates p21 for transcription (15). HES1 (Hairy and Enhancer of Split 1) is one of seven members of the HES family of basic helix-loop-helix (bHLH) transcription factors that is particularly well known as a repressive mediator of the canonical Notch signaling pathway (10). HES1 plays a key role in mediating Notch-dependent T cell lineage commitment (11), and has been reported to be an essential mediator of Notch-induced T cell acute lymphoblastic leukemia (T-ALL) (11,12). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (13). Transcription of cyclin D is in part regulated by the NICD binding to the promoter region of cyclin D (14). |
| 运输条件 | 0.75 |
| 存放说明 | -20C |
| 参考文献 | 1 . Li, X. and von Boehmer, H. (2011) ISRN Hematol 2011, 921706. 2 . Niimi, H. et al. (2007) J Cell Biol 176, 695-707. 3 . Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6. 4 . Wu, L. et al. (2002) Mol Cell Biol 22, 7688-700. 5 . Baudino, T.A. and Cleveland, J.L. (2001) Mol Cell Biol 21, 691-702. 6 . Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6. 7 . Lin, S.E. et al. (2002) J Biol Chem 277, 50612-20. 8 . Schroeter, E.H. et al. (1998) Nature 393, 382-6. 9 . Kitagawa, M. et al. (2001) Mol Cell Biol 21, 4337-46. 10 . Flores-Rozas, H. et al. (1994) Proc Natl Acad Sci U S A 91, 8655-9. 11 . Kobayashi, T. and Kageyama, R. (2010) Genes Cells 15, 689-98. 12 . Lukas, J. et al. (1996) Mol Cell Biol 16, 6917-25. 13 . Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35. 14 . Wendorff, A.A. et al. (2010) Immunity 33, 671-84. 15 . Espinosa, L. et al. (2010) Cancer Cell 18, 268-81. |
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition. | |
Western blot analysis of extracts from SK-N-MC, C6 and IMCD3 cells, using Cyclin D3 (DCS22) Mouse mAb. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Cyclin D3 (DCS22) Mouse mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Cyclin D3 (DCS22) Mouse mAb. | |
Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb. | |
Immunoprecipitation of p21 from human umbillical vein endothelial cells (HUVECs) using p21 Waf1/Cip1 (12D1) Rabbit mAb. Western blot detection was performed using the same antibody. | |
Confocal immunofluorescent analysis of MCF7 cells using p21 Waf1/Cip1 (12D1) Rabbit mAb (red) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using p21 Waf1/Cip1 (12D1) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Flow cytometric analysis of HeLa cells (red) and MCF7 cells (blue), using p21 Waf1/Cip1 (12D1) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded HeLa cells, transfected with SignalSilence® Control siRNA (Unconjugated) #6568 (left) or SignalSilence®p21 Waf1/Cip1 siRNA II #6558 (right), using p21 Waf1/Cip1 (12D1) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Notch1 (D1E11) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human inflammatory granulation tissue using Notch1 (D1E11) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded A2780 (left), Jurkat (center) and RL (right) cell pellets using Notch1 (D1E11) XP® Rabbit mAb. Both A2780 and Jurkat express Notch1, but only Jurkat cells have cleaved Notch1, while RL cells express very low or no Notch1. | |
Western blot analysis of total cell extract from various cell types using Notch1 (D1E11) XP® Rabbit mAb. The full-length (FL) Notch protein and the cleaved transmembrane/intracellular region (NTM) are indicated. | |
Western blot analysis of extracts from various cell lines using Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb (upper) or Notch1 (D1E11) XP® Rabbit mAb #3608 (lower). |
