| 货号 | 8357T |
| 描述 | The Stress and Apoptosis Antibody Sampler Kit provides an economical means of evaluating stress and apoptotic responses of each protein. The kit contains enough primary and secondary antibody to perform four western blot experiments per primary antibody. |
| 目标/特异性 | Each antibody in the Stress and Apoptosis Antibody Sampler Kit detects endogenous levels of target protein. Antibodies do not cross-react with any isoforms or phosphorylation sites of the target protein. |
| 供应商 | CST |
| 背景 | Cells respond to environmental or intracellular stresses through various mechanisms ranging from initiation of prosurvival strategies to activation of cell death pathways that remove damaged cells from the organism. Many of the proteins and cellular processes involved in normal signaling and survival pathways also play dual roles in cell death-promoting mechanisms. Apoptosis is a regulated cellular suicide mechanism characterized by nuclear condensation, cell shrinkage, membrane blebbing, and DNA fragmentation. Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). PARP appears to be involved in DNA repair in response to environmental stress (2). This protein can be cleaved by many ICE-like caspases in vitro (3,4) and is one of the main cleavage targets of caspase-3 in vivo (5,6). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (7). The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (8). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to a reduced interaction between p53 and its negative regulator, the oncoprotein MDM2 (9). MDM2 inhibits p53 accumulation by targeting it for ubiquitination and proteasomal degradation (10,11). Stress-activated protein kinases (SAPK)/Jun amino-terminal kinases (JNK) are members of the MAPK family that are activated by a variety of environmental stresses, inflammatory cytokines, growth factors, and GPCR agonists. Stress signals are delivered to this cascade by small GTPases of the Rho family (Rac, Rho, cdc42) (12). SAPK/JNK, when active as a dimer, can translocate to the nucleus and regulate transcription through its effects on c-Jun, ATF-2, and other transcription factors (12,13). c-Jun is a member of the Jun Family, containing c-Jun, JunB, and JunD, and is a component of the transcription factor AP-1 (activator protein-1). Extracellular signals from growth factors, chemokines, and stress activate AP-1-dependent transcription. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 through SAPK/JNK (reviewed in 14). AP-1 regulated genes exert diverse biological functions including cell proliferation, differentiation, and apoptosis, as well as transformation, invasion and metastasis, depending on cell type and context (13, 15-17). p38 MAP kinase (MAPK), also called RK (18) or CSBP (19), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (17-20). MKK3, MKK6, and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. MAPKAPK-2 is a direct target of p38 MAPK (17). Multiple residues of MAPKAPK-2 are phosphorylated in vivo in response to stress. However, only four residues (Thr25, Thr222, Ser272 and Thr334) are phosphorylated by p38 MAPK in an in vitro kinase assay (21). Phosphorylation at Thr222, Ser272, and Thr334 appears to be essential for the activity of MAPKAPK-2 (6). Heat shock protein (HSP) 27 is one of the small HSPs that are constitutively expressed at different levels in various cell types and tissues. In response to stress, the expression level of HSP27 increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78, and Ser82 by MAPKAPK-2 as a result of the activation of the p38 MAP kinase pathway (19,22). |
| 存放说明 | -20C |
| 参考文献 | 1 . Chehab, N.H. et al. (1999) Proc Natl Acad Sci U S A 96, 13777-82. 2 . Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4. 3 . Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358. 4 . Levine, A.J. (1997) Cell 88, 323-31. 5 . Rouse, J. et al. (1994) Cell 78, 1027-1037. 6 . Lazebnik, Y. A. et al. (1994) Nature 371, 346-347. 7 . Han, J. et al. (1994) Science 265, 808-11. 8 . Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803. 9 . Cohen, G.M. (1997) Biochem. J. 326, 1-16. 10 . Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69. 11 . Lee, J.C. et al. (1994) Nature 372, 739-46. 12 . Nicholson, D. W. et al. (1995) Nature 376, 37-43. 13 . Shieh, S.Y. et al. (1997) Cell 91, 325-34. 14 . Freshney, N.W. et al. (1994) Cell 78, 1039-49. 15 . Tewari, M. et al. (1995) Cell 81, 801-809. 16 . Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62. 17 . Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539. 18 . Honda, R. et al. (1997) FEBS Lett 420, 25-7. 19 . Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6. 20 . Davis, R.J. (2000) Cell 103, 239-52. 21 . Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3. 22 . Ben-Levy, R. et al. (1995) EMBO J. 14, 5920-5930. |
Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and either 10 μl of Phospho-c-Jun (Ser73) (D47G9) 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 SimpleChIP® using 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. | |
Immunohistochemical analysis of paraffin-embedded mouse lung using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb. | |
Western blot analysis of extracts from HeLa or HT-29 cells, untreated (-) or treated (+) with either UV (40 mJ/cm2 with 30 min recovery) or anisomycin (25 μg/mL, 30 min), using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb. | |
Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb. | |
Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or treated with λ phosphatase (middle), and NIH/3T3 cells (right) using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Negative staining in NIH/3T3 cells is in agreement with the observation that NIH/3T3 cells do not express HSP27 under basal conditions (5,7). | |
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or lambda phosphatase-treated (right), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb. | |
Immunohistochemical analysis of parafin-embedded human colon carcinoma using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb in the presence of control peptide (left) or Phospho-c-Jun (Ser73) Blocking Peptide (right). | |
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or UV-treated (right), using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb. | |
Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr), Jurkat cells, untreated or etoposide-treated (25 μM, overnight), and THP-1 cells, untreated or cycloheximide-treated (CHX, 10 μg/ml, overnight) followed by treatment with hTNF-α #8902 (20 ng/ml, 4 hr), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625 (upper), or total PARP Antibody #9542 (lower). Western blot 检测细胞提取物，分别取自未处理 (-)或Staurosporine #9953 (1 μM, 3 hr)处理的HeLa细胞，未处理 (-)或依托泊甙(25 μM, 过夜)处理的Jurkat细胞，未处理 (-)或放线菌酮(CHX, 10 μg/ml,过夜)处理然后用TNF-α #8902 (20 ng/ml, 4 hr)处理的THP-1细胞，使用抗体为Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb 兔单抗#5625 （上图）或PARP Antibody 多抗#9542（下图）。 | |
Western blot analysis of extracts from C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine (1 µM, 3 hrs) or etoposide (25 µM, 5 hrs) as indicated, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664. Western blot分析未处理或星胞菌素 (1uM, 3hrs)处理或依托泊苷(25uM, 5hrs)处理的C6 (大鼠源)、NIH/3T3 (小鼠源)和Jurkat (人源) 细胞的细胞提取物，抗体采用Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 兔单抗#9664. |
