| 货号 | 8333T |
| 目标/特异性 | Each antibody in the Fos Family Antibody Sampler Kit recognizes endogenous levels of the specific target protein. FosB (5G4) Rabbit mAb detects both FosB and FosB2 isoforms. Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb recognizes endogenous levels of FRA1 protein only when phosphorylated at Ser265. This antibody may also cross-react with phospho-FRA2, but does not cross-react with phospho-c-Fos or phospho-FosB. |
| 供应商 | CST |
| 背景 | The Fos family of nuclear oncogenes includes c-Fos, FosB, Fos-related antigen 1 (FRA1), and Fos-related antigen 2 (FRA2) (1). While most Fos proteins exist as a single isoform, the FosB protein exists as two isoforms: full-length FosB and a shorter form, FosB2 (Delta FosB), that lacks the carboxy-terminal 101 amino acids (1-3). The expression of Fos proteins is rapidly and transiently induced by a variety of extracellular stimuli including growth factors, cytokines, neurotransmitters, polypeptide hormones, and stress. Fos proteins dimerize with Jun proteins (c-Jun, JunB, and JunD) to form Activator Protein-1 (AP-1), a transcription factor that binds to TRE/AP-1 elements and activates transcription. Fos and Jun proteins contain the leucine-zipper motif that mediates dimerization and an adjacent basic domain that binds to DNA. The various Fos/Jun heterodimers differ in their ability to transactivate AP-1 dependent genes. In addition to increased expression, phosphorylation of Fos proteins by Erk kinases in response to extracellular stimuli may further increase transcriptional activity (4-6). Phosphorylation of c-Fos at Ser32 and Thr232 by Erk5 increases protein stability and nuclear localization (5). Phosphorylation of FRA1 at Ser252 and Ser265 by Erk1/2 increases protein stability and leads to overexpression of FRA1 in cancer cells (6). Following growth factor stimulation, expression of FosB and c-Fos in quiescent fibroblasts is immediate, but very short-lived, with protein levels dissipating after several hours (7). FRA1 and FRA2 expression persists longer and appreciable levels can be detected in asynchronously growing cells (8). Deregulated expression of c-Fos, FosB, or FRA2 can result in neoplastic cellular transformation; however, Delta FosB lacks the ability to transform cells (2,3). |
| 存放说明 | -20C |
| 参考文献 | Tulchinsky, E. (2000) Histol Histopathol 15, 921-8. Dobrazanski, P. et al. (1991) Mol Cell Biol 11, 5470-8. Nakabeppu, Y. and Nathans, D. (1991) Cell 64, 751-9. Rosenberger, S.F. et al. (1999) J Biol Chem 274, 1124-30. Sasaki, T. et al. (2006) Mol Cell 24, 63-75. Basbous, J. et al. (2007) Mol Cell Biol 27, 3936-50. Kovary, K. and Bravo, R. (1991) Mol Cell Biol 11, 2451-9. Kovary, K. and Bravo, R. (1992) Mol Cell Biol 12, 5015-23. |
Western blot analysis of extracts from HeLa cells, serum-starved overnight, and either left untreated or treated with TPA #4174 for 4 hours, using Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb #5841 (upper) and FRA1 (D80B4) Rabbit mAb #5281 (lower). Antibody phospho-specificity is shown by treating lysates with λ phosphatase. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with β-NGF #5221 (50 ng/ml) for 2 hr and either 10 μl of FosB (5G4) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and either 10 μl of Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with Human β-Nerve Gowth Factor (hβ-NGF) #5221 (50ng/ml) for 2h and either 10 μl of Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from serum-starved HeLa cells, untreated (-) or treated (+) with TPA #4174 (4 hrs), using FRA1 (D80B4) Rabbit mAb #5281 (upper) and β-Actin Antibody #4967 (lower).Western blot方法检测细胞提取物：未经处理(-)和TPA #4174 (4 hrs)处理(+)的无血清培养HeLa细胞。使用的抗体是FRA1 (D80B4) Rabbit mAb #5281(上图) 和β-Actin Antibody #4967 (下图)。 | |
Western blot analysis of extracts from HeLa and H-4-II-E cells serum-starved overnight and treated with TPA #4174 (4 hrs), using c-Fos (9F6) Rabbit mAb #2250.western blot方法检测细胞提取物：无血清培养过夜、TPA #4174 (4 hrs)处理的HeLa、H-4-II-E细胞。使用的抗体是c-Fos (9F6) Rabbit mAb #2250。 | |
Western blot analysis of extracts from HeLa cells serum-starved overnight and treated with TPA #4174 (4 hrs), or NIH/3T3 cells and C6 cells serum-starved overnight and serum-stimulated (4 hrs), using FosB (5G4) Rabbit mAb #2251.western blot方法检测细胞提取物：无血清培养过夜、TPA #4174 (4 hrs)处理的HeLa细胞，以及NIH/3T3细胞和无血清培养过夜和血清刺激(4 hrs)的C6细胞。使用的抗体是FosB (5G4) Rabbit mAb #2251。 | |
Western blot analysis of extracts from HeLa cells, serum-starved overnight and then either untreated or treated with TPA #4174 (4 hrs), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb #5348 (upper) or c-Fos (9F6) Rabbit mAb #2250 (lower). Antibody phospho-specificity is shown by treating lysates with λ phosphatase.Western blot方法检测细胞提取物：未经处理(-)和TPA #4174 (4 hrs)处理(+)的无血清培养过夜HeLa细胞。使用的抗体是Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb #5348(上图) 和c-Fos (9F6) Rabbit mAb #2250 (下图)。 | |
Western blot analysis of extracts from HeLa cells, serum-starved overnight, and either left untreated or treated with TPA #4174 for 4 hours, using Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb #5841 (upper) and FRA1 (D80B4) Rabbit mAb #5281 (lower). Antibody phospho-specificity is shown by treating lysates with λ phosphatase. | |
Western blot analysis of extracts from serum-starved HeLa cells, untreated or stimulated with TPA #4174 for 4 hours, using FRA1 (D80B4) Rabbit mAb (upper) and β-Actin Antibody #4967 (lower). | |
Western blot analysis of extracts from HeLa cells, serum-starved overnight and then either untreated or stimulated for 4 hours with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174, using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb (upper) and c-Fos (9F6) Rabbit mAb #2250 (lower). Antibody phospho-specificity is shown by treating lysates with λ-phosphatase. | |
Flow cytometric analysis of HeLa cells, untreated (blue) or TPA-treated (green), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb. | |
Confocal immunofluorescent analysis of HeLa cells, serum-starved (left), TPA-treated (middle) or treated with TPA and λ-phosphatase (right), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). | |
Confocal immunofluorescent analysis of HeLa cells either serum-starved (left) or TPA-treated (right) and labeled with FosB (5G4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). | |
Flow cytometric analysis of HeLa cells, untreated (blue) or TPA treated (green), using FosB (5G4) Rabbit mAb compared to a nonspecific negative control antibody (red). |
