Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb #5348

Cellular oncogene fos   fos   G0/G1 switch regulatory protein 7 (G0S7)  

No. Size Price
5348S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价
5348T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
5348 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 62 Rabbit IgG
IP 1:100
F 1:800
IF-IC 1:800
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Hamster, Monkey, Bovine, Pig, Horse,

Specificity / Sensitivity

Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb detects endogenous levels of c-Fos protein only when phosphorylated on Ser32. The antibody does not cross-react with other Fos proteins, including FosB, FRA1 and FRA2.

Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb兔单抗能检测内源性Ser32位点磷酸化的总c-Fos蛋白水平。该抗体不能与其他Fos蛋白包括FosB、 FRA1 、FRA2发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to Ser32 of human c-Fos protein.

该单克隆抗体是通过用与人源c-Fos蛋白Ser32位点周围氨基酸相一致的磷酸化肽段免疫动物后获得的。

IF-IC

IF-IC

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).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or TPA-treated (green), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb.

Western Blotting

Western Blotting

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.

Chromatin IP

Chromatin IP

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.

Background

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).

细胞核原癌基因的Fos家族成员包括c-Fos、FosB、Fos-相关抗原1 (FRA1)、Fos-相关抗原2 (FRA2)。虽然大部分Fos蛋白以单一亚型存在,但是FosB蛋白以两种亚型存在:全长的FosB蛋白和缩短的形式——FosB2 (Delta FosB),后者缺乏羧端的101个氨基酸(1-3)。Fos蛋白的表达是由细胞外刺激子快速且瞬间诱导的,这些刺激因子包括生长因子、细胞因子、神经传导物质、多肽激素和应激反应。Fos蛋白和Jun蛋白(c-Jun, JunB, 和JunD)结合成二聚体形成活化因子蛋白-1 (AP-1),它是结合到TRE/AP-1元件上的转录因子,并且能激活转录。Fos、Jun蛋白包含一个亮氨酸拉链基序,能调节二聚化和结合DNA的一个毗邻碱性域。各种Fos/Jun二聚体的不同点在于反式激活AP-1依赖性基因的能力。Fos蛋白对细胞外刺激因子应答后,除了表达量增加,同时也被Erk蛋白磷酸化,它的磷酸化可能会进一步增强转录活性(4-6)。Erk5蛋白引起的c-Fos蛋白丝氨酸(32位点)和苏氨酸(232位点)磷酸化增强了蛋白质稳定性和核定位(5)。Erk1/2引起的FRA1蛋白丝氨酸(252、265位点)磷酸化增强了蛋白稳定性,导致癌细胞内FRA1蛋白的过量表达(6)。生长因子刺激后,相对静止的成纤维细胞中FosB、c-Fos蛋白表达在短时间内接近蛋白水平,但是几个小时后就消失了(7)。FRA1、 FRA2的表达持续时间更长,而且在异步生长细胞中能检测到它的相对水平(8)。c-Fos, FosB, 和FRA2的失控表达将导致细胞的癌变;然而Delta FosB蛋白能阻止这种细胞转化的能力(2,3)。

  1. Tulchinsky, E. (2000) Histol Histopathol 15, 921-8.
  2. Dobrazanski, P. et al. (1991) Mol Cell Biol 11, 5470-8.
  3. Nakabeppu, Y. and Nathans, D. (1991) Cell 64, 751-9.
  4. Rosenberger, S.F. et al. (1999) J Biol Chem 274, 1124-30.
  5. Sasaki, T. et al. (2006) Mol Cell 24, 63-75.
  6. Basbous, J. et al. (2007) Mol Cell Biol 27, 3936-50.
  7. Kovary, K. and Bravo, R. (1991) Mol Cell Biol 11, 2451-9.
  8. Kovary, K. and Bravo, R. (1992) Mol Cell Biol 12, 5015-23.

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